Extended, high dose VEGF antagonist regimens for treatment of angiogenic eye disorders

Abstract

To provide optimal methods for treating or preventing angiogenic eye disorders and the like by administering a VEGF antagonist.SOLUTION: The present invention provides methods comprising administering one or more doses of about 8 mg or more of aflibercept once every 24 weeks to an eye of a subject with an angiogenic eye disorder, neovascular age related macular degeneration, diabetic retinopathy and / or diabetic macular edema; or comprising administering a single initial dose of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more secondary doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of the VEGF receptor fusion protein, in which each secondary dose is administered about 2 to 4 weeks after the immediately preceding dose and each tertiary dose is administered about 24 weeks after the immediately preceding dose.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Sequence Listing Reference This application incorporates by reference a computer readable sequence listing in ST.26 XML format entitled 11554WO01_Sequence, created May 10, 2024, and containing 5,467 bytes.

[0002] The field of the invention relates to methods of treating or preventing intraocular neovascular diseases by administering a VEGF antagonist. [Background technology]

[0003] The PHOTON clinical trial in DME and the PULSAR clinical trial in wAMD are double-blind, active-controlled pivotal trials conducted at multiple centers worldwide. In both trials, patients were randomized to one of three treatment arms to receive either aflibercept 8 mg every 12 weeks, aflibercept 8 mg every 16 weeks, or EYLEA every 8 weeks.

[0004] In both trials, patients treated with aflibercept 8 mg received three monthly initial doses, while patients treated with Eylea received five monthly initial doses in PHOTON and three monthly initial doses in PULSAR. During the first year, patients in the aflibercept 8 mg group had their dosing interval reduced to every 8 weeks if protocol-defined disease progression criteria were observed. The interval could not be extended until the second year of the trial. All Eylea patients maintained their fixed 8-week dosing regimen throughout their clinical trial participation. Summary of the Invention

[0005] The present invention relates to intraocular neovascular diseases (e.g., age-related macular degeneration (neovascularized (nAMD)), macular edema (ME), macular edema after retinal vein occlusion (ME-RVO), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), diabetic macular edema (DME), choroidal neovascularization (CNV), iris neovascularization, neovascular glaucoma, postoperative fibrosis in glaucoma, proliferative vitreoretinopathy (PVR), optic disc neovascularization, corneal neovascularization, and the like). 1. A method of treating or preventing neovascularization, retinal neovascularization, vitreous neovascularization, pannus, pterygium, vascular retinopathy, diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (e.g., characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53) or proliferative diabetic retinopathy, e.g., in a subject not suffering from DME), and / or diabetic retinopathy in a subject with diabetic macular edema (DME), , one or more doses of aflibercept (e.g., ≥ 8 mg) are administered, and the clearance of free aflibercept from the ocular compartment after intravitreal injection of aflibercept is about 0.367 to 0.458 mL / day (e.g., 0.41 mL / day), and the time for the amount of free aflibercept in the subject's ocular compartment after said intravitreal injection of aflibercept to reach the lower limit of quantitation (LLOQ) is about 15 weeks, and the time for the amount of free aflibercept in the subject's ocular compartment to reach the lower limit of quantitation (LLOQ) is about 15 weeks. the method includes administering the aflibercept such that the time for free aflibercept to reach the lower limit of quantitation (LLOQ) (e.g., about 0.0156 mg / L) in the subject's plasma after the intravitreal injection is about 3.5 weeks, wherein the aflibercept is administered in an aqueous pharmaceutical formulation, and the aflibercept contains less than about 3.5% high-molecular-weight molecular species immediately after production and purification, and / or contains about 6% or less high-molecular-weight molecular species after storage at about 2 to 8°C for about 24 months.In one embodiment of the present invention, the aqueous pharmaceutical formulation comprises at least about 100 mg / ml of a VEGF receptor fusion protein comprising two polypeptides, each comprising a second immunoglobulin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, and a multimerization component; about 10 to 100 mM L-arginine, sucrose, a histidine-based buffer, and a surfactant; wherein the formulation has a pH of about 5.0 to about 6.8; and the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification, and / or about 6% or less high molecular weight species after storage at about 2 to 8°C for about 24 months. In one embodiment of the invention, the method comprises administering a single initial dose of at least about 8 mg of aflibercept, followed by one or more secondary doses of at least about 8 mg of aflibercept, followed by one or more tertiary doses of at least about 8 mg of aflibercept, where each secondary dose is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose, and where each tertiary dose is administered about 24 weeks after the immediately preceding dose.

[0006] The present invention provides a method for slowing the clearance of free aflibercept from an ocular compartment following intravitreal injection compared to the clearance rate of aflibercept from the ocular compartment following intravitreal injection of ≦4 mg of aflibercept, comprising intravitreously injecting into an eye of a subject in need thereof a single initial dose of at least about 8 mg of aflibercept, followed by one or more secondary doses of at least about 8 mg of aflibercept, followed by one or more tertiary doses of at least about 8 mg of aflibercept, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose. In one embodiment of the invention, the clearance of free aflibercept from the ocular compartment is about 34% slower than the clearance from the ocular compartment after intravitreal injection of ≦4 mg aflibercept, e.g., the clearance of free aflibercept from the ocular compartment is about 0.367-0.458 mL / day or 0.41 mL / day after intravitreal injection of ≧8 mg aflibercept.

[0007] The present invention also provides a method of increasing the time for the amount of free aflibercept in a subject's ocular compartment to reach the lower limit of quantitation (LLOQ) after intravitreal injection of aflibercept, compared to the time for the amount of free aflibercept in the subject's ocular compartment to reach the LLOQ after intravitreal injection of about 2 mg of aflibercept, e.g., by more than 1.3 weeks, e.g., between about 6 weeks and more than 10 weeks, e.g., to about 15 weeks, comprising intravitreously injecting into an eye of a subject in need thereof a single initial administration of at least about 8 mg of aflibercept, followed by one or more secondary administrations of at least about 8 mg of aflibercept, followed by one or more tertiary administrations of at least about 8 mg of aflibercept, wherein each secondary administration is administered about 2 to 4 weeks after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration.

[0008] The present invention also provides a method of increasing the time to reach the lower limit of quantitation (LLOQ) (e.g., about 0.0156 mg / L) in a subject's plasma after intravitreal injection of aflibercept, compared to the time to reach the LLOQ in the subject's plasma after intravitreal injection of about 2 mg of aflibercept, e.g., by more than 1.5 weeks, for example, by about 2 to 3.5 weeks, comprising intravitreously injecting into an eye of a subject in need thereof a single initial administration of at least about 8 mg of aflibercept, followed by one or more secondary administrations of at least about 8 mg of aflibercept, followed by one or more tertiary administrations of at least about 8 mg of aflibercept, wherein each secondary administration is administered about 2 to 4 weeks after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration. In one embodiment of the present invention, ≧8 mg of aflibercept is administered in an aqueous pharmaceutical formulation comprising aflibercept, the aqueous pharmaceutical formulation comprising one or more of a histidine-based buffer, arginine (e.g., L-arginine such as L-arginine hydrochloride), a sugar such as sucrose, or a polyol, and having a pH of about 5.8. In one embodiment of the present invention, aflibercept has less than about 3.5% high molecular weight species immediately after production and purification, and / or has about 6% or less high molecular weight species after about 24 months of storage at about 2 to 8°C, for example, ≥ 8 mg of aflibercept is in an aqueous pharmaceutical formulation comprising at least about 100 mg / ml of a VEGF receptor fusion protein, the VEGF receptor fusion protein comprising two polypeptides each comprising a second immunoglobulin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, and a multimerization component, and about 10 to 100 mM L-arginine, sucrose, a histidine-based buffer, and a surfactant, wherein the formulation has a pH of about 5.0 to about 6.8, and the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification, and / or has about 6% or less high molecular weight species after about 24 months of storage at about 2 to 8°C.

[0009] The present invention relates to intraocular neovascular diseases (e.g., age-related macular degeneration (nAMD)), macular edema (ME), macular edema after retinal vein occlusion (ME-RVO), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), diabetic macular edema (DME), choroidal neovascularization (CNV), iris neovascularization, neovascular glaucoma, postoperative fibrosis in glaucoma, proliferative vitreoretinopathy (PVR), optic nerve head neovascularization, corneal neovascularization, retinal neovascularization, vitreous neovascularization, pannus, pterygium, vascular retinopathy, diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53) or proliferative diabetic retinopathy, e.g., and / or diabetic macular edema (DME) in a subject with nAMD, DR, and / or DME in need of treatment or prevention (e.g., diabetic retinopathy in a subject not afflicted with DME, and / or diabetic macular edema (DME)), or for improving best corrected visual acuity (BCVA) in a subject with nAMD, DR, and / or DME in need of treatment or prevention, or for promoting retinal dryness in a subject with nAMD, DR, and / or DME in need of treatment or prevention, the method comprising administering to the eye of the subject one or more doses of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, once every 24 weeks. In one embodiment of the invention, the method comprises administering to the subject's eye a single initial dose of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, followed by one or more secondary doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of the VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks (preferably 4 weeks) after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose.

[0010] The present invention relates to intraocular neovascular diseases (e.g., age-related macular degeneration (nAMD)), macular edema (ME), macular edema after retinal vein occlusion (ME-RVO), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), diabetic macular edema (DME), choroidal neovascularization (CNV), iris neovascularization, neovascular glaucoma, postoperative fibrosis in glaucoma, proliferative vitreoretinopathy (PVR), optic nerve head neovascularization, corneal neovascularization, retinal neovascularization, vitreous neovascularization, pannus, pterygium, vascular retinopathy, diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53), or proliferative diabetic retinopathy (DR) (e.g., diabetic retinopathy characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53). and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to the eye of the subject a single initial dose of about 8 mg or more of a VEGF-receptor fusion protein, preferably aflibercept, followed by one or more secondary doses of about 8 mg or more of the VEGF-receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of the VEGF-receptor fusion protein, wherein each secondary dose is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose.

[0011] In an embodiment of the present invention, a subject is diagnosed with an intraocular neovascular disease (e.g., age-related macular degeneration (neovascularized (nAMD)), macular edema (ME), macular edema after retinal vein occlusion (ME-RVO), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), diabetic macular edema (DME), choroidal neovascularization (CNV), iris neovascularization, neovascular glaucoma, postoperative fibrosis in glaucoma, proliferative vitreoretinopathy (PVR), optic disc neovascularization, corneal neovascularization, retinal neovascularization, vitreous neovascularization, pannus, pterygium, vascular retinopathy, diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (e.g., diabetic retinopathy that is about 47 or 53 years old)). A method for treating or preventing diabetic retinopathy (e.g., diabetic retinopathy characterized by a Diabetic Retinopathy Severity Scale (DRSS) level) or proliferative diabetic retinopathy (e.g., in subjects not afflicted with DME), and / or diabetic retinopathy in subjects with diabetic macular edema (DME) comprises administering to a subject in need thereof 8 mg of a VEGF receptor fusion protein (0.07 mL or 70 microliters), preferably aflibercept, via intravitreal injection every four weeks (approximately every 28 days + / - 7 days, monthly) for the first three doses, and then once every 24 weeks (6 months + / - 7 days).

[0012] The present invention also relates to intraocular neovascular diseases (e.g., age-related macular degeneration (neovascularized (nAMD)), macular edema (ME), macular edema after retinal vein occlusion (ME-RVO), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), diabetic macular edema (DME), choroidal neovascularization (CNV), iris neovascularization, neovascular glaucoma, postoperative fibrosis in glaucoma, proliferative vitreoretinopathy (PVR), optic disc neovascularization, corneal neovascularization, retinal neovascularization, vitreous neovascularization, pannus, pterygium, vascular retinopathy, diabetic retinopathy (DR) ( For example, a method of treating or preventing non-proliferative diabetic retinopathy (e.g., characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53) or proliferative diabetic retinopathy, e.g., in a subject not afflicted with DME, and / or diabetic retinopathy in a subject with diabetic macular edema (DME) in a subject in need thereof, comprising: (1) administering to the subject an initial dose of 2 mg of a VEGF receptor fusion protein; and then, one month later, administering to the subject an initial dose of 8 mg of a VEGF receptor fusion protein. (1) the subject is administered an initial dose of 2 mg of VEGF receptor fusion protein, and then one month later, the method comprises administering to the subject a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month thereafter by ... second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter, one or more maintenance doses of 8 mg of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (2) the subject is administered an initial dose of 2 mg of VEGF receptor fusion protein, and then one month later, the method comprises administering to the subject a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month thereafter by a second 8 mg secondary dose of VEGF receptor fusion protein. (3) the subject is administered an initial dose of 2 mg of VEGF receptor fusion protein, then one month later, the method comprises administering to the subject a second 8 mg dose of VEGF receptor fusion protein, followed by one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 24 weeks thereafter according to an HDq24 dosing regimen; or(4) the subject is administered an initial dose of 2 mg of VEGF receptor fusion protein, then one month later, the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein and an additional 8 mg maintenance dose of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; or (5) the subject is administered an initial dose of 2 mg of VEGF receptor fusion protein and one month later, a first dose of 2 mg of VEGF receptor fusion protein, then one more month later, the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein and an additional 8 mg maintenance dose of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen. (5) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (6) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and then the method (6) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (7) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and then the method further comprises administering to the subject one month later a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen. or (8) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein and, one month later, a first 2 mg secondary dose of VEGF receptor fusion protein, and then, one more month later, the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein and, one month later, every 24 weeks according to the HDq24 dosing regimen.or (9) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, one month later a first 2 mg secondary dose of VEGF receptor fusion protein, and one month later a second 2 mg secondary dose of VEGF receptor fusion protein, and then one month later the method comprises administering to the subject an initial dose of 8 mg of VEGF receptor fusion protein, one month thereafter a first 8 mg secondary dose of VEGF receptor fusion protein, one month thereafter a second 8 mg secondary dose of VEGF receptor fusion protein, and then once every 24 weeks thereafter according to an HDq24 dosing regimen. or (10) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and one more month later by a second 2 mg secondary dose of VEGF receptor fusion protein, and then one more month later, the method comprises administering to the subject a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; (11) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and another month later by a second 2 mg secondary dose of VEGF receptor fusion protein, and then the method comprises administering to the subject another 1 month later a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (12) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen. (13) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, and another 2 mg secondary dose of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein two months later, and all of the additional 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; and then one month later, the subject receives an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by a HDq24 dose regimen. or (14) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, and then one month later by administering to the subject a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later byor (14) a subject has received an initial 2 mg dose of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed by a second 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 8 mg secondary dose of VEGF receptor fusion protein, followed by a second 8 mg secondary dose of VEGF receptor fusion protein, followed by a second 8 mg secondary dose of VEGF receptor fusion protein, followed by a third 8 mg secondary dose of VEGF receptor fusion protein, followed by a second 8 mg secondary dose of VEGF receptor fusion protein, followed by a third 8 mg secondary dose of VEGF receptor fusion protein, followed by a fourth ... The method includes administering to the subject, one month later, a second 8 mg secondary dose of VEGF receptor fusion protein, followed by one or more 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks thereafter according to an HDq24 dosing regimen; or (16) the subject administers an initial 2 mg dose of VEGF receptor fusion protein, one month later, a first 2 mg secondary dose of VEGF receptor fusion protein, another month later, a second 2 mg secondary dose of VEGF receptor fusion protein, and another month later, a third 2 mg secondary dose of VEGF receptor fusion protein. and then, two months later, the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein, followed by an additional 8 mg maintenance dose of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; or (17) the subject receives an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed another month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed another month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein. (17) the subject has been administered an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, and then two months later, the method comprises administering to the subject an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (18) the subject has been administered an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, and then two months later, the method comprises administering to the subject an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen;and administering an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed another month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, and then two months later, administering to the subject a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein. (19) a subject receives an initial 2 mg dose of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed by a fourth 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, followed by a fourth 2 mg secondary dose of VEGF receptor fusion protein, followed by a fourth 2 mg secondary dose of VEGF receptor fusion protein, followed by a fifth 2 mg secondary dose of VEGF receptor fusion protein, followed by a fifth 2 mg secondary dose of VEGF receptor fusion protein, followed by a sixth ... (20) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, and then two months later, the method comprises administering to the subject a second 8 mg secondary dose of VEGF receptor fusion protein, and then over the next 24 weeks, one or more 24-week 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (21) the subject has been administered an initial dose of 2 mg of VEGF receptor fusion protein followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein. and a second 2 mg secondary dose of VEGF receptor fusion protein one month later, a third 2 mg secondary dose of VEGF receptor fusion protein one month later, and a fourth 2 mg secondary dose of VEGF receptor fusion protein one month thereafter, and then the method comprises administering to the subject two months later a first 8 mg maintenance dose of VEGF receptor fusion protein, and all further 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; (21) A subject is administered an initial dose of 2 mg of a VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of the VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of the VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of the VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of the VEGF receptor fusion protein, and one or more 2 mg maintenance doses every eight weeks thereafter; then the method comprises administering to the subject, two months after the last maintenance dose of the VEGF receptor fusion protein, a second 2 mg secondary dose of the VEGF receptor fusion protein, followed by one or more 2 mg maintenance doses every eight weeks thereafter. (21) a subject receives an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then every 24 weeks thereafter by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (22) a subject receives an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein. and then, two months after the last VEGF receptor fusion protein maintenance dose, the subject receives a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, and one or more 2 mg maintenance doses every eight weeks thereafter; and then, two months after the last VEGF receptor fusion protein maintenance dose, the subject receives a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, followed once every 24 weeks thereafter according to the HDq24 dosing regimen. or (23) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, a first 2 mg secondary dose of VEGF receptor fusion protein one month later, a second 2 mg secondary dose of VEGF receptor fusion protein one month later, a third 2 mg secondary dose of VEGF receptor fusion protein one month later, a fourth 2 mg secondary dose of VEGF receptor fusion protein one month later, and one or more 2 mg maintenance doses every eight weeks thereafter, and then the method comprises:(24) the subject receives an initial dose of 2 mg of VEGF receptor fusion protein, followed one month later by a first 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a third 2 mg secondary dose of VEGF receptor fusion protein, followed one month later by a fourth 2 mg secondary dose of VEGF receptor fusion protein, followed by one or more 2 mg maintenance doses every eight weeks thereafter. and then, two months after the last maintenance dose of VEGF receptor fusion protein, the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein, followed by all additional 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen, the HDq24 dosing regimen comprising a single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more secondary doses of about 8 mg or more of VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose and each tertiary dose is administered about 24 weeks after the immediately preceding dose.

[0013] The present invention also provides a method of treating or preventing neovascular age-related macular degeneration (nAMD), diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof and receiving a dosing regimen for treating or preventing said disorders, comprising: (a) administering to the subject an initial dose of 8 mg of a VEGF receptor fusion protein, then one month later, administering to the subject a first 8 mg secondary dose of the VEGF receptor fusion protein, and one month thereafter administering a second 8 mg secondary dose of the VEGF receptor fusion protein; and then every 24 weeks thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen; or (b) the subject has been administered an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, and then after another month, the method comprises administering to the subject a second 8 mg secondary dose of VEGF receptor fusion protein, followed by one or more 8 mg maintenance doses of VEGF receptor fusion protein according to an HDq24 dosing regimen every 24 weeks thereafter. or (c) the subject has been administered an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, and another month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then 24 weeks later the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein, and all additional 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; or (d) the subject has been administered an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and then 24 weeks later the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein, followed by all additional 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; The method comprises administering an initial dose of 8 mg of VEGF receptor fusion protein, followed one month later by a first 8 mg secondary dose of VEGF receptor fusion protein, followed another month later by a second 8 mg secondary dose of VEGF receptor fusion protein, and every 24 weeks thereafter, the subject receives one or more 8 mg maintenance doses of VEGF receptor fusion protein, and then 24 weeks after the last maintenance dose of VEGF receptor fusion protein, the method comprises administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 24 weeks according to the HDq24 dosing regimen.and administering a further 8 mg maintenance dose of VEGF receptor fusion protein to all of the patients, wherein the HDq24 dosing regimen comprises a single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more secondary doses of about 8 mg or more of VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose.

[0014] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD), diabetic retinopathy, or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye a single initial administration of about 8 mg or more of a VEGF-receptor fusion protein, followed by one or more secondary administrations of about 8 mg or more of the VEGF-receptor fusion protein, followed by one or more tertiary administrations of about 8 mg or more of the VEGF-receptor fusion protein, wherein each secondary administration is administered about 2 to 4 weeks after the immediately preceding administration, and each tertiary administration is The method further comprises administering the tertiary administration about 8 weeks, 12 weeks, 16 weeks, or 20 weeks after the immediately preceding administration, and further comprising, if the tertiary administration at 8, 12, 16, or 20 weeks has expired, evaluating the patient for suitability to extend the interval between tertiary administrations, and increasing the interval between tertiary administrations in 4-week increments if, in the judgment of the treating physician based on the patient's visual acuity and / or central retinal thickness, it is appropriate to extend the interval between tertiary administrations, e.g., the interval is extended up to about 24 weeks, e.g., after one or more 4-week increases in the interval.

[0015] The present invention also provides a method for treating or preventing an intraocular neovascular disease (preferably nAMD, DR, and / or DME) in a subject in need thereof, the subject being treated with a dosing regimen for treating or preventing the disorder, comprising a single initial dose of about 2 mg of a VEGF receptor fusion protein, preferably aflibercept, followed by one or more secondary doses of about 2 mg of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 2 mg of the VEGF receptor fusion protein, wherein each secondary dose is administered in a dose that is about 2 mg of the immediately preceding dose. and each third administration is administered about 8 weeks after the immediately preceding administration, wherein the subject is on a 2 mg VEGF receptor fusion protein dosing regimen, the method comprising administering an 8 mg dose of VEGF receptor fusion protein to the subject's eye, evaluating the subject about 4, 8, 10, or 12 weeks after administration, and if, in the treating physician's judgment, dosing every 24 weeks is appropriate, continuing to administer 8 mg of VEGF receptor fusion protein to the subject every 24 weeks thereafter.

[0016] In one embodiment of the invention, the subject of the methods of the invention is receiving a single initial dose of about 2 mg of a VEGF receptor fusion protein, preferably aflibercept, followed by two, three, or four secondary doses of about 2 mg of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 2 mg of the VEGF receptor fusion protein, for treating or preventing neovascularized age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema, where each secondary dose is administered about 4 weeks after the immediately preceding dose, and each tertiary dose is administered about 8 weeks after the immediately preceding dose.

[0017] The present invention also provides a method for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, or diabetic macular edema in a subject in need thereof, comprising administering to the eye of the subject a single initial administration of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, followed by one or more secondary administrations of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary administrations of about 8 mg or more of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding administration, and each tertiary administration is administered about 12 or 16 weeks after the immediately preceding administration, and further comprising extending the interval between tertiary administrations from 12, 16, or 20 weeks to 24 weeks after the immediately preceding administration after receiving one or more of the tertiary administrations about 12 or 16 weeks after the immediately preceding administration. For example, in one embodiment of the present invention, during the aforementioned treatment, the subject exhibits (a) a loss of <5 letters in BCVA, and / or (b) a central retinal thickness (CRT) of <300 or 320 μm. In embodiments of the present invention, the method further comprises assessing BVCA and / or CRT in the subject, and extending the interval between tertiary doses if the subject (a) exhibits a loss of <5 letters in BCVA, and / or (b) exhibits a CRT of <300 or 320 μm.

[0018] The present invention also provides a method for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye a single initial dose of about ≥8 mg of a VEGF receptor fusion protein, preferably aflibercept, followed by one or more secondary doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of the VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks (preferably 4 weeks) after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose, further comprising shortening the interval between the tertiary doses from 24 to 12, 16, or 20 after receiving one or more of the tertiary doses about 24 weeks after the immediately preceding dose. In one embodiment of the present invention, during the treatment, the subject demonstrates (a) a loss of >10 letters in BCVA compared to baseline, and / or (b) an increase in CRT of >50 μm compared to baseline. For example, in an embodiment of the invention, the method further comprises assessing BVCA and / or CRT in the subject, and shortening the interval between tertiary administrations if the subject (a) exhibits a loss of >10 letters of BCVA relative to baseline, and / or (b) exhibits an increase in CRT of >50 μm relative to baseline.

[0019] In one embodiment of the present invention, (a) A loss of more than 5 letters of BCVA (ETDRS) compared with the BCVA observed approximately 12 weeks after the start of treatment; (b) an increase in CRT of more than 25 micrometers is observed compared to the CRT observed at about 12 weeks after the start of treatment; and / or (c) New onset foveal neovascularization or foveal hemorrhage For example, if it is 16 or 20 weeks after initiation of treatment, the interval between the third doses may be reduced from 24 weeks to 12, 16, or 20 weeks; or (a) A loss of more than 5 letters of BCVA (ETDRS) compared with the BCVA observed approximately 12 weeks after the start of treatment; (b) an increase in CRT of more than 25 micrometers is observed compared to the CRT observed at about 12 weeks after the start of treatment; and / or (c) New onset foveal neovascularization or foveal hemorrhage For example, if it is the 24th week after initiation of treatment, the interval between the third doses is shortened from 24 weeks to 12 weeks, 16 weeks, or 20 weeks.

[0020] The present invention provides a method for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye three doses of about 8 mg of a VEGF-receptor fusion protein, preferably aflibercept, in a formulation containing about 114.3 mg / ml of the VEGF-receptor fusion protein, once every four weeks, and after the three doses, administering one or more doses of the VEGF-receptor fusion protein at extended intervals up to 24 weeks.

[0021] The present invention provides a method for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye a single initial administration of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, followed by two secondary administrations of about 8 mg or more of a VEGF receptor fusion protein, each secondary administration being administered about 2-4 weeks (preferably 4 weeks) after the immediately preceding administration, followed by one or more tertiary administrations every 12, 16, 20, or 24 weeks; and, after administration, determining whether the subject meets at least one criterion for shortening or lengthening the interval between the one or more tertiary administrations of the VEGF receptor fusion protein by 2 weeks, 3 weeks, 4 weeks, or 2-4 weeks. and b) if a determination is made, administering additional doses of the VEGF receptor fusion protein with a shortened or extended interval between doses, wherein criteria for extending the interval include 1. loss of <5 letters of BCVA, and / or 2. CRT <300 or 320 micrometers, and criteria for shortening the interval include 1. loss of >10 letters of BCVA; 2. persistent or worsening DME, and / or 3. increase in CRT >50 micrometers. In one embodiment of the present invention, criteria for extending the interval include both 1. loss of <5 letters of BCVA from week 12, and 2. CRT <300 or 320 micrometers as measured by SD-OCT; and / or criteria for shortening the interval include both 1. loss of >10 letters of BCVA, e.g., associated with persistent or worsening DME, from week 12, and 2. increase in CRT >50 micrometers, e.g., from week 12. In one embodiment of the present invention, if the criteria are met, the interval is extended to 24 weeks.

[0022] The present invention provides a method for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, the subject having been previously treated with one or more 2 mg doses of a VEGF-receptor fusion protein, preferably aflibercept, the method comprising administering to the subject's eye a single initial administration of at least about 8 mg of a VEGF-receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF-receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF-receptor fusion protein, wherein each secondary administration is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration.

[0023] In one embodiment of the present invention, subjects with certain exclusion criteria are excluded from treatment, or are not excluded from treatment if the exclusion criteria are not met. For example, a subject with any one or more of an ocular or periorbital infection, active intraocular inflammation, and / or hypersensitivity is excluded from ocular administration of a VEGF receptor fusion protein. In one embodiment of the present invention, the method further comprises evaluating the subject for an ocular or periorbital infection, active intraocular inflammation, and / or hypersensitivity, and excluding the subject from administration if any one or more of the following conditions are present in the subject:

[0024] In one embodiment of the present invention, the subject is monitored for adverse events such as conjunctival hemorrhage, cataract, vitreous detachment, vitreous floaters, corneal epithelial defects, and / or elevated intraocular pressure, etc. If such an AE is identified, the identified AE may be treated and / or such treatment or prophylaxis may be discontinued.

[0025] In one embodiment of the present invention, the method involves pre-administration preparation of a VEGF receptor fusion protein, preferably aflibercept. For example, the method may include providing or having a glass vial containing approximately 70 microliters of an aqueous formulation comprising 8 mg of a VEGF receptor fusion protein, the glass vial having a protective plastic cap and stopper, packaged together with an inventive needle, e.g., an 18 gauge x 1.5 inch, 5 micron filter needle including a tip and a bevel, a 30 gauge x 0.5 inch syringe needle, and a syringe, e.g., a 1-mL Luer Lock syringe with a graduation mark marking the 70 microliter volume, before each administration; then (1) visually inspecting the aqueous formulation in the vial and, if particulate, cloudy, or discolored, using another vial of aqueous formulation containing a VEGF receptor fusion protein; (2) removing the protective plastic cap from the vial; and (3) cleaning the top of the vial with an alcohol wipe, followed by aseptic technique. (4) removing an 18 gauge x 1.5 inch, 5 micron filter needle and a 1 mL syringe from their packaging; (5) attaching the filter needle to the syringe by twisting the filter needle onto the tip of a Luer-lock syringe; (6) pushing the filter needle into the center of the vial stopper until the filter needle is fully inserted into the vial and the tip touches the bottom or bottom edge of the vial; (7) holding the vial in an upright position and tilting it slightly while ensuring the bevel of the filter needle is submerged in the liquid, withdrawing all of the contents of the VEGF receptor fusion protein vial into the syringe; (8) continuing to tilt the vial during withdrawal, keeping the bevel of the filter needle submerged in the formulation; (9) retracting the plunger rod sufficiently as the vial is emptied to completely empty the filter needle; (10) removing the filter needle from the syringe and discarding it; and (11) inserting a 30 gauge x 0.(11) removing a 5-inch needle from its packaging and twisting the needle onto the tip of a Luer-lock syringe to securely attach the needle to the syringe; (12) holding the syringe with the needle pointing up, checking for air bubbles in the syringe and, if any, gently flicking the syringe with a finger until the bubbles rise to the top; and (13) slowly pushing the plunger so that the plunger tip is aligned with the graduation mark marking the 70 microliter mark on the syringe. In one embodiment of the present invention, the injection of the VEGF receptor fusion protein is performed under controlled aseptic conditions, including surgical hand hygiene and the use of sterile gloves, sterile drapes, and a sterile eye speculum (or equivalent), and anesthesia and topical broad-spectrum antibacterial eye drops are administered prior to injection.

[0026] In one embodiment of the present invention, a subject is receiving a dosing regimen for treating or preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema requiring a single initial dose of about 2 mg of VEGF receptor fusion protein, followed by two, three, or four secondary doses of about 2 mg of VEGF receptor fusion protein, followed by one or more tertiary doses of about 2 mg of VEGF receptor fusion protein, wherein each secondary dose is administered about 4 weeks after the immediately preceding dose, and each tertiary dose is administered about 8 weeks after the immediately preceding dose, and the subject is at any stage (initial, secondary, or tertiary) of the 2 mg VEGF receptor fusion protein dosing regimen.

[0027] In one embodiment of the invention, 24 weeks is 6 months, 168 days, or twice a year, and the one or more secondary administrations is two secondary administrations, and 2-4 weeks is about 4 weeks, and 12-20 weeks is about 12 weeks, and 12-20 weeks is about 16 weeks, and 12-20 weeks is about 20 weeks, and 12-20 weeks is about 12-16 weeks, and 8-16 weeks is about 12 weeks, and 8-16 weeks is about 16 weeks, and 8-16 weeks is about 12-16 weeks, and 2-4 weeks is about 4 weeks, and the one or more secondary administrations is two secondary administrations, and 12-20 weeks is about 12 weeks, and the one or more secondary administrations is two secondary administrations, and 12-20 weeks is about 16 weeks, and the one or more secondary administrations is two secondary administrations, and 12-20 weeks is about 20 weeks, and the one or more secondary administrations is two secondary administrations, and 12-20 weeks is about 12-16 weeks, wherein the one or more secondary doses are two secondary doses, wherein 2 to 4 weeks is about 4 weeks, wherein the one or more secondary doses are two secondary doses, and the VEGF receptor fusion protein is aflibercept, wherein 12 to 20 weeks is about 12 weeks, wherein the one or more secondary doses are two secondary doses, and the VEGF receptor fusion protein is aflibercept, wherein 12 to 20 weeks is about 16 weeks, wherein the one or more secondary doses are two secondary doses, and the VEGF receptor fusion protein is aflibercept, wherein 12 to 20 weeks is about 20 weeks, wherein the one or more secondary doses are two secondary doses, and the VEGF receptor fusion protein is aflibercept, and / or wherein 12 to 20 weeks is about 12 to 16 weeks, wherein the one or more secondary doses are two secondary doses, and the VEGF receptor fusion protein is aflibercept.

[0028] In one embodiment of the present invention, the VEGF receptor fusion protein comprises amino acids 27 to 457 of the amino acid sequence set forth in SEQ ID NO: 2 and is selected from the group consisting of aflibercept and conbercept, and comprises two polypeptides comprising: (1) a VEGFR1 component comprising amino acids 27 to 129 of SEQ ID NO: 2, (2) a VEGFR2 component comprising amino acids 130 to 231 of SEQ ID NO: 2, and (3) a multimerization component comprising amino acids 232 to 457 of SEQ ID NO: 2; comprises two polypeptides comprising a second immunoglobulin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, and a multimerization component; comprises two polypeptides comprising a second immunoglobulin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, a fourth Ig domain of VEGFR2, and a multimerization component; or comprises two VEGFR1R2-Fc ΔC1(a) polypeptides encoded by the nucleic acid sequence of SEQ ID NO: 1. In one embodiment of the present invention, the VEGF receptor fusion protein comprises or consists of amino acids 27 to 457 of the amino acid sequence set forth in SEQ ID NO:2.

[0029] In one embodiment of the invention, the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation selected from the group consisting of A-KKKK. In one embodiment of the invention, the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 114.3 mg / ml of the VEGF receptor fusion protein, preferably aflibercept.

[0030] In one embodiment of the present invention, the VEGF receptor fusion protein is administered to both eyes of a subject.

[0031] In one embodiment of the present invention, the VEGF receptor fusion protein, preferably aflibercept, is administered from a syringe or pre-filled syringe, e.g., made of glass or plastic and / or sterile, using, e.g., a 30 gauge x 0.5 inch sterile needle.

[0032] In one embodiment of the invention, the subject has previously received one or more doses of a VEGF receptor fusion protein, such as, for example, 2 mg of Eylea. One or more additional doses other than those specifically mentioned may also be administered to the subject.

[0033] In one embodiment of the present invention, a subject receiving 2 mg of a VEGF receptor fusion protein, preferably aflibercept, receives the protein in an aqueous pharmaceutical formulation containing 40 mg / ml of the VEGF receptor fusion protein, 10 mM sodium phosphate, 40 mM NaCl, 0.03% polysorbate 20, and 5% sucrose, at a pH of 6.2.

[0034] In one embodiment of the present invention, 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising ≥100 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine (preferably L-arginine), e.g., a sugar or polyol (e.g., sucrose). In one embodiment of the present invention, the formulation has a pH of about 5.8. For example, the formulation may comprise about 103-126 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine, and in one embodiment, about 114.3 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0035] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is delivered in a volume of about 100 μl or less, about 75 μl or less, about 70 μl or less, or about 50 μl, 51 μl, 52 μl, 53 μl, 54 μl, 55 μl, 56 μl, 57 μl, 58 μl, 59 μl, 60 μl, 61 μl, 62 μl, 63 μl, 64 μl, 65 μl, 66 μl, 67 μl, 68 μl, 69 μl, 70 μl, 71 μl, 72 μl, 73 μl, 74 μl, 75 μl, 76 μl, 77 μl, 78 μl, 79 μl, 80 μl, 81 μl, 82 μl, 83 μl, 84 μl, 85 μl, 86 μl, 87 μl, 88 μl, 89 μl, 90 μl, 91 μl, 92 μl, 93 μl, 94 μl, 95 μl, 96 μl, 97 μl, 98 μl, 99 μl, 100 μl, 101 μl, 102 μl, 103 μl, 104 μl, 105 μl, 106 μl, 107 μl, 108 μl, 109 μl, 110 μl, 111 μl, 112 μl, 113 μl, 114 μl, 115 μl, 116 μl, 117 μl, 118 μl, 119 μl, 120 μl, 121 μl, 12 It may be administered in a volume of 3 μl, 74 μl, 75 μl, 76 μl, 77 μl, 78 μl, 79 μl, 80 μl, 81 μl, 82 μl, 83 μl, 84 μl, 85 μl, 86 μl, 87 μl, 88 μl, 89 μl, 90 μl, 91 μl, 92 μl, 93 μl, 94 μl, 95 μl, 96 μl, 97 μl, 98 μl, 99 μl, or 100 μl, for example, in a volume of about 70±4 or 5 microliters.

[0036] In one embodiment of the present invention, the methods herein comprise administering a VEGF receptor fusion protein, preferably aflibercept, to both eyes of a subject, e.g., intravitreally.

[0037] In one embodiment of the present invention, the subject has an improvement in Diabetic Retinopathy Severity Scale (DRSS), an improvement in best corrected visual acuity, dry retina, an increase in best corrected visual acuity, a BCVA of at least 69 letters, a fluid-free fovea, a decrease in central retinal thickness (CRT), no vascular leakage as measured by fluorescein angiography (FA), and a National Eye Institute Visual Function Questionnaire (NEI-VFQ). Improvement from pre-treatment baseline in the total score of the ETDRS Questionnaire, fluid-free retina in the fovea and central subfield (total fluid, intraretinal fluid [IRF] and / or subretinal fluid [SRF]), maintenance of fluid-free retina (total fluid, intraretinal fluid [IRF] and / or subretinal fluid [SRF] in the fovea and central subfield), loss of macular edema, fluid-free retina on spectral-domain optical coherence tomography (SD-OCT), no deviation from HDq12 or HDq16 or HDq20 treatment regimen once initiated, administered intravitreally at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, then approximately once every 8 weeks or once every 2 months, non-inferior BVCA compared with aflibercept, an increase in BCVA (according to the ETDRS letter score) of approximately 7, 8, or 9 letters by 60 weeks from the start of treatment, where baseline BCVA is approximately 61, 62, or 63 letters. BCVA (according to ETDRS letter score) of at least about 69 letters by 48 or 60 weeks after initiation of treatment; no loss of 5, 10, 15, or 69 letters of BCVA after 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment; improvement in BCVA (according to ETDRS letter score) by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment; an improvement in BVCA by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks from initiation; a BCVA score (according to ETDRS letter score) of about 69, 70, 71, 72, or 73 between 48 and 60 weeks; and a change in BCVA score (according to ETDRS letter score) from initiation of treatment of about 7, 8, or 9 between 36 and 48 weeks.A BCVA of approximately 60 or 70 at any time point between 36 and 48 weeks; a change in BCVA score (according to the ETDRS letter score) from the start of treatment of approximately 7, 8, or 9 between 48 and 60 weeks, with a BCVA of approximately 69, 70, 71, 72, or 73 at any time point between 48 and 60 weeks; A gain of ≥ 4 letters, ≥ 5 letters, ≥ 6 letters, ≥ 7 letters, ≥ 8 letters, > 9 letters, or > 10 letters in BCVA as measured by the ETDRS eye chart or equivalent Snellen visual acuity by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks from the start of treatment; no loss of 5, 10, or 15 letters (according to ETDRS letter score) by 48 or 60 weeks from the start of treatment; a gain of at least 5, 10, or 15 letters (according to ETDRS letter score) by 48 or 60 weeks from the start of treatment; an improvement in BCVA of approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) by 4 weeks after the start of treatment if receiving an HDq12 regimen, or approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; and a loss of ≥ 4 letters (ETDRS or equivalent Snellen visual acuity) by 8 weeks after the start of treatment if receiving an HDq12 regimen. an improvement in BCVA of approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 12 weeks after initiation of treatment; an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 16 weeks after initiation of treatment; an improvement in BCVA of approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 20 weeks after initiation of treatmentor an improvement in BCVA of approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, and an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 24 weeks after initiation of treatment; and an improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 28 weeks after initiation of treatment. By 32 weeks after initiation of treatment, an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 36 weeks after initiation of treatment, an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 or 7 letters if receiving an HDq16 regimen Improvement in BCVA by approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 40 weeks after initiation of treatment; improvement in BCVA by approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 44 weeks after initiation of treatment an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 48 weeks after initiation of treatment; an improvement in BCVA by approximately 8 weeks after initiation of treatment that is maintained through at least 48 weeks thereafter during the treatment regimen; and an improvement in BCVA of approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, by 4 weeks after initiation of treatmentor a BCVA of approximately 66 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 8 weeks after starting treatment, a BCVA of approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or a BCVA of approximately 67 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 12 weeks after starting treatment, a BCVA of approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or a BCVA of approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen By 16 weeks after the start of treatment, a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 71 letters if receiving an HDq12 regimen or a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 69 letters if receiving an HDq16 regimen; by 20 weeks after the start of treatment, a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 70 letters if receiving an HDq12 regimen or a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 68 letters if receiving an HDq16 regimen; By 28 weeks after treatment initiation, patients on the HDq12 regimen had a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 72 letters if they were on the HDq12 regimen, or approximately 70 letters if they were on the HDq16 regimen. By 32 weeks after treatment initiation, patients on the HDq12 regimen had a BCVA (ETDRS or equivalent Snellen visual acuity) of approximately 72 letters if they were on the HDq12 regimen, or approximately 70 letters if they were on the HDq16 regimen. 1 letter BCVA (ETDRS or equivalent Snellen visual acuity), or approximately 70 letters of BCVA (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; approximately 71 letters of BCVA (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 68 letters of BCVA (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 36 weeks after initiation of treatment; approximately 72 letters of BCVA (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, by 40 weeks after initiation of treatmentor a BCVA of approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, and a BCVA of approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or a BCVA of approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 44 weeks after initiation of treatment; By 48 weeks after starting treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 48 weeks after starting treatment, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen and baseline BCVA of approximately <73 ETDRS letters; by 48 weeks after starting treatment, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen and baseline BVA of approximately <73 ETDRS letters An improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if A is approximately >73 ETDRS letters; an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if baseline BCVA is approximately <73 ETDRS letters by 48 weeks after initiation of treatment if receiving an HDq16 regimen; an improvement in BCVA of approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if baseline BCVA is approximately >73 ETDRS letters by 48 weeks after initiation of treatment if receiving an HDq16 regimen; By 48 weeks after initiation of treatment, an improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if the baseline CRT is approximately < 400 micrometers and the patient is receiving an HDq12 regimen; an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) if the baseline CRT is approximately > 400 micrometers and the patient is receiving an HDq12 regimen; and an improvement in BCVA of approximately < 400 micrometers and the patient is receiving an HDq16 regimen by 48 weeks after initiation of treatment. an improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if the baseline CRT is approximately >400 micrometers by 48 weeks after initiation of treatment, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) if the baseline CRT is approximately >400 micrometers by 48 weeks after initiation of treatment if receiving an HDq16 regimen, a gain of >5, >10, or ≥15 letters of BCVA (according to ETDRS letter score) by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment≥2 or >3 step improvement in the Diabetic Retinopathy Severity Scale (DRSS) by 90 or 96 weeks; ≥2 step improvement in the Diabetic Retinopathy Severity Scale (DRSS) by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks from the start of treatment; ≥100% fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the fovea and central subfield by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks from the start of treatment, as measured by optical coherence tomography (OCT). free retina; no vascular leakage as measured by fluorescein angiography (FA) from the start of treatment through 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks; maintenance of a fluid-free retina (total fluid in the fovea and central subfields, IRF, and / or SRF) from the start of treatment through 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks; total area of ​​fluorescein leakage within the ETDRS grid (mm2) at 48 or 60 weeks measured by fluorescein angiography of approximately 12, 13, or 14 mm; 2a reduction of ≥ 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks from the start of treatment, a fluid-free retina by spectral-domain optical coherence tomography (SD-OCT), a fluid-free retina (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the fovea by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks from the start of treatment, a dry retina by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks from the start of treatment, as measured by optical coherence tomography (OCT), a fluid-free fovea by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks; a change in central retinal thickness by 4 weeks after initiation of treatment of approximately -118 or -118.3 micrometers if receiving an HDq12 regimen, or approximately -124 or -125 or -124.9 or -125.5 micrometers if receiving an HDq16 regimen; and a change in central retinal thickness by 8 weeks after initiation of treatment of approximately -137 or -137.4 micrometers if receiving an HDq12 regimen, or approximately -140 or -140.9 or -140.5 micrometers if receiving an HDq16 regimen. a change in central retinal thickness of about -139 or -140 or -139.6 or -140.3 micrometers if receiving an HDq12 regimen, by 12 weeks after initiation of treatment, or about -150 or -150.1 micrometers if receiving an HDq12 regimen, or about -152 or -153 or -152.7 or -153.4 micrometers if receiving an HDq16 regimen; a change in central retinal thickness of about -139 or -139.4 micrometers if receiving an HDq12 regimen, by 16 weeks after initiation of treatment, or about -150 or -150.1 micrometers if receiving an HDq12 regimen, or about -152 or -153 or -152.7 or -153.4 micrometers if receiving an HDq16 regimen, by 16 weeks after initiation of treatment A change in central retinal thickness of approximately -145 or -146 or -145.5 or -146.4 micrometers if receiving a HDq16 regimen, by 20 weeks after initiation of treatment, and approximately -117 or -117.1 micrometers if receiving a HDq12 regimen, or approximately -112 or -113 or -112.5 or -113.3 micrometers if receiving a HDq16 regimen, or approximately -158 or -158 if receiving a HDq12 regimen, by 24 weeks after initiation of treatment.A change in central retinal thickness of approximately -103 or -104 or -103.8 or -104.3 micrometers if receiving an HDq16 regimen by 28 weeks after initiation of treatment; a change in central retinal thickness of approximately -146 or -147 or -146.7 micrometers if receiving an HDq12 regimen, or approximately -162 or -162.3 micrometers if receiving an HDq16 regimen by 32 weeks after initiation of treatment; and a change in central retinal thickness of approximately -132 micrometers if receiving an HDq12 regimen by 32 weeks after initiation of treatment. a change in central retinal thickness of about -145 or -146 or -145.8 micrometers if receiving an HDq16 regimen, by 36 weeks after initiation of treatment; a change in central retinal thickness of about -168 or -168.1 micrometers if receiving an HDq12 regimen, or about -124 or -125 or -124.7 or -125.2 micrometers if receiving an HDq16 regimen, by 40 weeks after initiation of treatment; A change in central retinal thickness of approximately -122 or -123 or -122.5 or -123.1 micrometers if receiving an HDq16 regimen, by 44 weeks after initiation of treatment; a change in central retinal thickness of approximately -147 or -148 or -147.4 micrometers if receiving an HDq12 regimen, or approximately -164 or -164.1 or -164.3 micrometers if receiving an HDq16 regimen; and a change in central retinal thickness of approximately -171 or -172 or -173.1 micrometers if receiving an HDq12 regimen, by 48 weeks after initiation of treatment. A change in central retinal thickness of -171.7 micrometers, or about -148 or -149 or -148.3 or -149.4 micrometers if receiving an HDq16 regimen, by 60 weeks after initiation of treatment; a change in central retinal thickness of about -181.95 or -176.24 micrometers if receiving an HDq12 regimen, or about -166.26 or -167.18 micrometers if receiving an HDq16 regimen, and a decrease in central retinal thickness by 4, 5, 6, 7, or 8 weeks after initiation of treatment. This is maintained during the treatment regimen within a range of about ±17, ±18, or ±19 micrometers from the start of treatment through at least 48 weeks; a reduction in central retinal thickness by about 100, 125, 150, 175, or 200 micrometers from the start of treatment through 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks; a reduction in central retinal thickness of about 148 to 182 micrometers from the start of treatment through about 48 or 60 weeks, as measured by optical coherence tomography (OCT), compared to baseline. the central retinal thickness (CRT) is about 449, 450, 455, or 460 micrometers; a reduction in central retinal thickness (CRT) by at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 micrometers by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks from the start of treatment; and a plasma free aflibercept concentration of about 0.1667 days after the first dose. about 0.149 (±0.249) mg / L, where at baseline, the plasma free aflibercept concentration was undetectable and the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 0.3333 days after the first administration, the plasma free aflibercept concentration was about 0.205 (±0.250) mg / L, where at baseline, the plasma free aflibercept was undetectable and the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 1 day after the first administration, the plasma free aflibercept concentration was about 0.266 (±0.211) mg / L, where at baseline, the plasma free aflibercept was undetectable and the subject had not received intravitreal aflibercept treatment for at least 12 weeks; and about 2 days after the first administration, the plasma free aflibercept concentration was about 0.218 (±0.and about 4 days after the first dose, plasma free aflibercept is about 0.140 (±0.0741) mg / L, wherein at baseline, plasma free aflibercept is undetectable and the subject has not received intravitreal aflibercept treatment for at least 12 weeks. about 7 days after the first dose, the plasma free aflibercept concentration is about 0.0767 (±0.0436) mg / L, where at baseline there was no detectable free aflibercept in the plasma, and the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 14 days after the first dose, the plasma free aflibercept concentration is about 0.0309 (±0.0241) mg / L, where at baseline there was no detectable free aflibercept in the plasma, and the subject had not received intravitreal aflibercept treatment for at least 12 weeks; and about 21 days after the first dose, plasma free aflibercept is about 0.0171 (±0.0171) mg / L, whereby at baseline, there is no detectable free aflibercept in plasma, and the subject has not received intravitreal aflibercept treatment for at least 12 weeks. and about 28 days after the first dose, plasma free aflibercept is about 0.00730 (±0.0113 ... Free aflibercept in plasma is undetectable, the subject has not received intravitreal aflibercept treatment for at least 12 weeks, and about 0.1667 days after the first dose, adjusted bound aflibercept in plasma is about 0.00698 (±0.0276) mg / L, and at baseline, adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks, and about 0.and after 3333 days, the plasma adjusted bound aflibercept is about 0.00731 (±0.0279) mg / L, where at baseline, the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; and about 1 day after the first dose, the plasma adjusted bound aflibercept is about 0.0678 (±0.0486) mg / L, where at baseline, the adjusted bound aflibercept is about and about 2 days after the first dose, the plasma adjusted bound aflibercept is about 0.138 (±0.0618) mg / L, where at baseline the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks. and about 4 days after the first dose, the plasma adjusted bound aflibercept is about 0.138 (±0.0618) mg / L, where at baseline the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks. and about 7 days after the first dose, the plasma adjusted bound aflibercept is about 0.346 (±0.151) mg / L, and at baseline, the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280). The subject has not received intravitreal aflibercept treatment for at least 12 weeks. The subject has about 7 days after the first dose, the plasma adjusted bound aflibercept is about 0.346 (±0.151) mg / L, and at baseline, the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280). and the subject had not received intravitreal aflibercept treatment for at least 12 weeks, and about 14 days after the first dose, the plasma adjusted bound aflibercept was about 0.374 (±0.110) mg / L, and at baseline, the adjusted bound aflibercept was about 0.00583 mg / L (±0.0280), and the subject had not received intravitreal aflibercept treatment for at least 12 weeks, and about 21 days after the first dose, the plasma adjusted bound aflibercept was about 0.343 (±0.and about 28 days after the first dose, the plasma adjusted bound aflibercept is about 0.269 (±0.149) mg / L, wherein at baseline, the adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks. The subjects had not received intravitreal aflibercept treatment for at least 12 weeks; the peak plasma concentration of free aflibercept was reached approximately 0.965 days after the first dose; the peak plasma concentration of free aflibercept was reached to be approximately 0.310 mg / L (±0.263); the free aflibercept in plasma was approximately 0 to approximately 1.08 mg / L; and the peak plasma concentration (mg / L) of free aflibercept per mg dose of aflibercept was approximately 0.0388 (±0.0328) mg / L / mg. Achieving and / or maintaining one or more of the following about 14 days after the first dose: reaching a peak plasma concentration of adjusted conjugated aflibercept; reaching a peak plasma concentration of adjusted conjugated aflibercept of about 0.387 mg / L (±0.135); plasma adjusted conjugated aflibercept of about 0.137 to about 0.774 mg / L; a peak plasma adjusted conjugated aflibercept (mg / L) per aflibercept dose (mg) of about 0.0483 (±0.0168) mg / L / mg; absence of anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment; improvement from pre-treatment baseline in the National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score; and / or loss of macular edema. For example, in one embodiment of the present invention, dry retina is characterized by a loss of intraretinal and / or subretinal fluid, or dry retina is characterized by the absence of intraretinal fluid (IRF) and subretinal fluid (SRF) in a subject's eye after the subject receives three monthly doses of a VEGF receptor fusion protein, preferably aflibercept.

[0038] In an embodiment of the invention, a subject will experience an improvement in BCVA of about 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or about 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen by 64 weeks after initiation of treatment; an improvement in BCVA of about 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or about 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen by 68 weeks after initiation of treatment; an improvement in BCVA of about 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or about 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen by 72 weeks after initiation of treatment. An improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 76 weeks after initiation of treatment; an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen, by 80 weeks after initiation of treatment; ) or approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 84 weeks after initiation of treatment, an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 88 weeks after initiation of treatment, an improvement in BCVA of approximately 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen by 92 weeks after initiation of treatment, an improvement in BCVA of approximately 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 96 weeks after initiation of treatment, an improvement in BCVA of approximately 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 64 weeks after initiation of treatment,A BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 68 weeks after initiation of treatment, a BCVA of approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 72 weeks after initiation of treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen by 76 weeks after initiation of treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or a BCVA of approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 80 weeks after initiation of treatment, a BCVA of approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or a BCVA of approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen By 84 weeks after initiation of treatment, a BCVA of approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 88 weeks after initiation of treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen By 92 weeks after initiation of treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 96 weeks after initiation of treatment, a BCVA of approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen; by 64 weeks after initiation of treatment,A change in central retinal thickness of approximately -173.4 micrometers or approximately -173 micrometers if receiving an HDq12 regimen, or approximately -164.3 micrometers or approximately -164 micrometers if receiving an HDq16 regimen, by 68 weeks after starting treatment; a change in central retinal thickness of approximately -159.4 micrometers or approximately -159 micrometers if receiving an HDq12 regimen, or approximately -153.9 micrometers or approximately -154 micrometers if receiving an HDq16 regimen, by 72 weeks after starting treatment , a change in central retinal thickness of approximately -166.6 micrometers or approximately -167 micrometers if receiving an HDq12 regimen, or approximately -134.2 micrometers or approximately -134 micrometers if receiving an HDq16 regimen, by 76 weeks after starting treatment; and a change in central retinal thickness of approximately -181.1 micrometers or approximately -181 micrometers if receiving an HDq12 regimen, or approximately -160.8 micrometers or approximately -161 micrometers if receiving an HDq16 regimen, by 80 weeks after starting treatment. , a change in central retinal thickness of about -168.9 micrometers or about -169 micrometers if receiving an HDq12 regimen, or about -164 micrometers or about -164 micrometers if receiving an HDq16 regimen, by 84 weeks after initiation of treatment; a change in central retinal thickness of about -177.5 micrometers or about -178 micrometers if receiving an HDq12 regimen, or about -150.2 micrometers or about -150 micrometers if receiving an HDq16 regimen, by 88 weeks after initiation of treatment; A change in central retinal thickness of approximately -171.2 micrometers or approximately -171 micrometers if receiving an HDq12 regimen, or approximately -144.3 micrometers or approximately -144 micrometers if receiving an HDq16 regimen, by 92 weeks after initiation of treatment; a change in central retinal thickness of approximately -166.7 micrometers or approximately -167 micrometers if receiving an HDq12 regimen, or approximately -155.5 micrometers or approximately -156 micrometers if receiving an HDq16 regimen, by 96 weeks after initiation of treatmentA change in central retinal thickness of about -185.3 micrometers or about -185 micrometers if receiving an HDq12 regimen, or about -155 micrometers or about -155 micrometers if receiving an HDq16 regimen; by 64 weeks after initiation of treatment, a central retinal thickness of about 279.4 micrometers if receiving an HDq12 regimen, or about 289.6 micrometers if receiving an HDq16 regimen; by 68 weeks after initiation of treatment, a central retinal thickness of about 294.5 micrometers if receiving an HDq12 regimen, or about 305.3 micrometers if receiving an HDq16 regimen; by 72 weeks after initiation of treatment, a central retinal thickness of about 284.2 micrometers if receiving an HDq12 regimen, or about 327.2 micrometers if receiving an HDq16 regimen; by 76 weeks after initiation of treatment, a central retinal thickness of about 270.6 micrometers if receiving an HDq12 regimen, or about 302 micrometers if receiving an HDq16 regimen By 80 weeks after initiation of treatment, central retinal thickness of approximately 284.6 micrometers for those receiving an HDq12 regimen or approximately 293.5 micrometers for those receiving an HDq16 regimen; by 84 weeks after initiation of treatment, central retinal thickness of approximately 274.7 micrometers for those receiving an HDq12 regimen or approximately 310.8 micrometers for those receiving an HDq16 regimen; by 88 weeks after initiation of treatment, central retinal thickness of approximately 283.7 micrometers for those receiving an HDq12 regimen or approximately 293.5 micrometers for those receiving an HDq16 regimen; Achieve and / or maintain one or more of the following by 92 weeks after initiation of treatment: a central retinal thickness of about 312.3 micrometers if receiving an HDq12 regimen, or about 301.8 micrometers if receiving an HDq16 regimen; and by 96 weeks after initiation of treatment: a central retinal thickness of about 267.5 micrometers if receiving an HDq12 regimen, or about 304.2 micrometers if receiving an HDq16 regimen.

[0039] In one embodiment of the invention, references to 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60 weeks from the start of treatment are about 48 weeks or 60 weeks from the start of treatment.

[0040] In one embodiment of the present invention, a subject is administered one initial dose, two secondary doses, and three tertiary doses of a VEGF receptor fusion protein, e.g., aflibercept, in the first year; or one initial dose, two secondary doses, and two tertiary doses of a VEGF receptor fusion protein, e.g., aflibercept, in the first year; or one initial dose, two secondary doses, and three tertiary doses of a VEGF receptor fusion protein, e.g., aflibercept, in the first year, followed by two to four tertiary doses in the second year.

[0041] In one embodiment of the present invention, the interval between doses is adjusted (increased / maintained / decreased) based on visual and / or anatomical outcomes, for example according to the criteria set forth in FIG. 3 and / or FIG.

[0042] The present invention also provides a kit comprising a container comprising a VEGF receptor fusion protein and instructions for use of the VEGF receptor fusion protein, wherein the container is a vial or a prefilled syringe, the container comprises ≥ 100 mg / mL of the VEGF receptor fusion protein, the container comprises ≥ 114.3 mg / mL of the VEGF receptor fusion protein, the instructions comprise instructions for administering aflibercept to a patient with DR, DME and / or nAMD, the instructions comprise instructions that aflibercept 8 mg treatment is to be initiated with three consecutive doses, one injection per month (once every four weeks), the instructions comprise instructions that after the initial three consecutive doses, the injection interval may be extended up to every 24 weeks, and the instructions comprise instructions that the treatment interval may be adjusted based on a physician's judgment of the visual and / or anatomical outcome.

[0043] The present invention provides aflibercept for use in treating or preventing intraocular neovascular disease, neovascularized age-related macular degeneration, diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the treatment or prevention comprising administering aflibercept in one or more doses at an interval and in an amount such that clearance of free aflibercept from the ocular compartment after intravitreal injection of aflibercept is about 0.367 to 0.458 mL / day, wherein the time for the amount of free aflibercept in the subject's ocular compartment to reach the lower limit of quantitation (LLOQ) after the intravitreal injection of aflibercept is about 15 weeks, and the time for free aflibercept in the subject's plasma to reach the lower limit of quantitation (LLOQ) after the intravitreal injection of aflibercept is about 3.5 weeks.

[0044] The present invention provides aflibercept for use in a method for slowing the clearance of free aflibercept from an ocular compartment following intravitreal injection compared to the clearance rate of aflibercept from the ocular compartment following intravitreal injection of <4 mg of aflibercept, the method comprising administering to an eye of a subject in need thereof a single initial dose of about 8 mg or more of aflibercept, followed by One or more second doses of aflibercept of about 8 mg or more, followed by one or more third doses of about 8 mg or more of aflibercept administered intravitreally, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose, Provided is aflibercept, wherein each third dose is administered about 24 weeks after the immediately preceding dose.

[0045] The present invention provides aflibercept for use in a method for increasing the time for the amount of free aflibercept in an ocular compartment of a subject to reach the lower limit of quantitation (LLOQ) after intravitreal injection of aflibercept, compared to the time for the amount of free aflibercept in the ocular compartment of the subject to reach the LLOQ after intravitreal injection of about 2 mg of aflibercept, comprising: The method comprises administering to an eye of a subject in need thereof: A single initial dose of approximately 8 mg or more of aflibercept, followed by One or more second doses of aflibercept of about 8 mg or more, followed by one or more third doses of about 8 mg or more of aflibercept administered intravitreally, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose, Provided is aflibercept, wherein each third dose is administered about 24 weeks after the immediately preceding dose.

[0046] The present invention provides aflibercept for use in a method for increasing the time for free aflibercept in the plasma of a subject to reach the lower limit of quantitation (LLOQ) after intravitreal injection of aflibercept, compared to the time for free aflibercept in the plasma of the subject to reach the LLOQ after intravitreal injection of about 2 mg of aflibercept, the method comprising intravitreously injecting into the eye of a subject in need thereof a single initial administration of at least about 8 mg of aflibercept, followed by one or more secondary administrations of at least about 8 mg of aflibercept, followed by one or more tertiary administrations of at least about 8 mg of aflibercept, wherein each secondary administration is administered about 2 to 4 weeks after the immediately preceding administration, and wherein each tertiary administration is administered about 24 weeks after the immediately preceding administration.

[0047] The present invention provides - for the treatment or prevention of intraocular neovascular disease, nAMD, diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, - To improve best-corrected visual acuity in a subject in need thereof who has intraocular neovascular disease, nAMD, DR, and / or DME; or 1. A VEGF receptor fusion protein for use in a method for promoting retinal dryness in a subject with retinal desiccation and / or DME in need thereof, comprising: The method includes administering to the subject's eye one or more doses of about 8 mg or more of the VEGF receptor fusion protein once every 24 weeks.

[0048] The present invention provides aflibercept for use in treating or preventing intraocular neovascular disease, neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein the treatment or prevention comprises three consecutive injections of 8 mg aflibercept, starting monthly (every four weeks), followed by one or more injections every 24 weeks, each of which has an aflibercept concentration of 114.3 mg / mL or a volume of 70 μL. In one embodiment of the present invention, the treatment interval between two subsequent 8 mg aflibercept injections is adjusted (increased / maintained / shortened) based on visual and / or anatomical outcomes, such as, but not limited to, gain or loss of a BCVA letter, increase or decrease in CRT, presence or absence of subretinal fluid, presence or absence of hemorrhage, or persistence or worsening of DME. In one embodiment of the present invention, the treatment interval is shortened by 2-4 weeks, 2 weeks, 3 weeks, or 4 weeks compared to the previous treatment interval if the subject is identified as meeting at least one of the following criteria for a reduction in treatment duration: 1. a loss of >5 or >10 letters in BCVA, a CRT of >300 or 320 μm, a decrease in CRT of >50 μm, or persistent or worsening DME. In one embodiment of the present invention, the treatment interval is extended by 2-4 weeks, 2 weeks, 3 weeks, or 4 weeks compared to the previous treatment interval if the subject is identified as meeting at least one of the following criteria for an extension of the treatment interval: a loss of <5 or <10 letters in BCVA, a CRT of <300 or 320 μm, a decrease in CRT of >50 μm, no subretinal fluid, or no hemorrhage.

[0049] The present invention provides a VEGF receptor fusion protein for use in treating or preventing an intraocular neovascular disease or diabetic macular edema in a subject in need thereof, the method comprising administering 8 mg of the VEGF receptor fusion protein (0.07 mL or 70 microliters) by intravitreal injection every four weeks (approximately every 28 days + / - 7 days, monthly) for the first three doses, followed by administering 8 mg of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 24 weeks (+ / - 7 days).

[0050] The present invention provides a VEGF receptor fusion protein for use in treating or preventing diabetic retinopathy (DR) in a subject in need thereof, the method comprising administering 8 mg of the VEGF receptor fusion protein (0.07 mL or 70 microliters) via intravitreal injection every four weeks (approximately every 28 days + / - 7 days, monthly) for the first three doses, followed by administering 8 mg of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 24 weeks (+ / - 7 days).

[0051] The present invention provides a VEGF receptor fusion protein for use in treating or preventing neovascular age-related macular degeneration in a subject in need thereof, the method comprising administering 8 mg of the VEGF receptor fusion protein (0.07 mL or 70 microliters) by intravitreal injection every four weeks (approximately every 28 days + / - 7 days, monthly) for the first three doses, followed by administering 8 mg of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 24 weeks (+ / - 7 days).

[0052] The present invention provides aflibercept for use in treating or preventing intraocular neovascular disease, neovascular age-related macular degeneration, diabetic macular edema, or diabetic retinopathy in a subject in need thereof, comprising administering to the subject's eye a single initial dose of 8 mg of aflibercept, followed by one or more tertiary doses of about 8 mg of aflibercept, each tertiary dose administered about 24 weeks after the immediately preceding dose. In one embodiment of the present invention, the subject is not treatment-naive, or the subject has been previously treated with a VEGF antagonist, or preferably, the subject has been previously treated with 8 mg aflibercept or 2 mg aflibercept.

[0053] The present invention provides aflibercept for use in the treatment or prevention of intraocular neovascular disease, neovascular age-related macular degeneration, diabetic macular edema, or diabetic retinopathy in a subject previously treated with 2 mg of aflibercept, comprising administering to the subject's eye a single initial dose of about 8 mg of aflibercept, followed by one or more secondary doses of about 8 mg of aflibercept, and then one or more tertiary doses of about 8 mg of aflibercept, wherein each secondary dose is administered about 4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose. In one embodiment of the present invention, the administration of one or more doses of 8 mg aflibercept to the subject's eye follows an HDq24 administration regimen or a Treat and Extent administration regimen.

[0054] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof and receiving a dosing regimen therefor, comprising: (a) the subject receives an initial dose of 8 mg of a VEGF receptor fusion protein, and then, one month later, the method comprises administering to the subject a first 8 mg secondary dose of a VEGF receptor fusion protein, followed one month later by a second 8 mg secondary dose of a VEGF receptor fusion protein, and then every 24 weeks thereafter by administering one or more 8 mg maintenance doses of a VEGF receptor fusion protein according to an HDq24 dosing regimen; or (b) the subject has received an initial dose of 8 mg of a VEGF receptor fusion protein and one month later a first 8 mg secondary dose of a VEGF receptor fusion protein, and then, one more month later, the method comprises administering to the subject a second 8 mg secondary dose of a VEGF receptor fusion protein, and then every 24 weeks thereafter one or more 8 mg maintenance doses of a VEGF receptor fusion protein according to an HDq24 dosing regimen; or (c) the subject receives an initial dose of 8 mg of a VEGF receptor fusion protein, and one month later a first 8 mg secondary dose of a VEGF receptor fusion protein, and another month later a second 8 mg secondary dose of a VEGF receptor fusion protein, and then 24 weeks later, the method comprises administering to the subject a first 8 mg maintenance dose of a VEGF receptor fusion protein, and all further 8 mg maintenance doses of a VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; or (d) the subject receives an initial dose of 8 mg of a VEGF receptor fusion protein, and one month later, a first 8 mg secondary dose of the VEGF receptor fusion protein, and another month later, a second 8 mg secondary dose of the VEGF receptor fusion protein, and every 24 weeks thereafter, the subject receives one or more 8 mg maintenance doses of the VEGF receptor fusion protein, and then 24 weeks after the final maintenance dose of the VEGF receptor fusion protein, administering to the subject one or more 8 mg maintenance doses of the VEGF receptor fusion protein, and all additional 8 mg maintenance doses of the VEGF receptor fusion protein every 24 weeks according to an HDq24 dosing regimen; The HDq24 dosing regimen comprises: A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by one or more secondary doses of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more third doses comprising about 8 mg or more of a VEGF receptor fusion protein; Each secondary dose is administered approximately 2 to 4 weeks after the previous dose, Each third administration is administered about 24 weeks after the immediately preceding administration, providing a VEGF receptor fusion protein.

[0055] The invention provides a VEGF receptor fusion protein for use in treating or preventing an intraocular neovascular disorder in a subject in need thereof, the subject being on a dosing regimen for treating or preventing the disorder requiring a single initial administration of about 2 mg of the VEGF receptor fusion protein, followed by one or more secondary administrations of about 2 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of about 2 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 4 weeks after the immediately preceding administration and each tertiary administration is administered about 8 weeks after the immediately preceding administration, the subject being on a 2 mg VEGF receptor fusion protein dosing regimen, the method comprising administering an 8 mg dose of the VEGF receptor fusion protein to the eye of the subject, evaluating the subject about 4, 8, 10, or 12 weeks after administration, and if, in the judgment of the treating physician, dosing every 24 weeks is appropriate, then continuing to administer to the subject 8 mg of the VEGF receptor fusion protein every 24 weeks.

[0056] The present invention provides a VEGF receptor fusion protein for use in treating or preventing intraocular neovascular disease, nAMD, diabetic retinopathy, or diabetic macular edema in a subject in need thereof, wherein the treatment or prevention comprises administering to the subject's eye a single initial administration of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more, preferably two, secondary administrations of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary administrations of about 8 mg or more of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2 to 4 weeks after the immediately preceding administration, and each tertiary administration is administered about 12, 16, or 20 weeks after the immediately preceding administration, and wherein one or more of the tertiary administrations are administered about 12, 16, or 20 weeks after the immediately preceding administration, and the interval between the tertiary administrations is about 12, 16, or 20 weeks after the immediately preceding administration. After 12 to 24 weeks, After 24 weeks from 26 weeks, or The VEGF receptor fusion protein further comprises an extension after 20 to 24 weeks.

[0057] The present invention provides a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular diseases, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye a single initial administration of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration and each tertiary administration is administered about 24 weeks after the immediately preceding administration, and wherein after one or more of the tertiary administrations are administered about 24 weeks after the immediately preceding administration, the interval between the tertiary administrations is: From 24 weeks to 8 weeks From 24 weeks to 12 weeks From 24 weeks to 16 weeks, or The present invention provides a VEGF receptor fusion protein, further comprising shortening the treatment period from 24 weeks to 20 weeks.

[0058] The present invention provides a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, comprising administering to the subject's eye three doses of about 8 mg of the VEGF receptor fusion protein, once every four weeks, in a formulation containing about 114.3 mg / ml of the VEGF receptor fusion protein, and, after the three doses, administering one or more doses of the VEGF receptor fusion protein at extended intervals of up to 24 weeks.

[0059] The present invention relates to a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular diseases, neovascularized age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, the VEGF receptor fusion protein comprising a single initial administration of about 8 mg or more of the VEGF receptor fusion protein, followed by two secondary administrations of about 8 mg or more of the VEGF receptor fusion protein, then administering one or more tertiary administrations of about 8 mg or more of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration and each tertiary administration is administered about 24 weeks after the immediately preceding administration, and after these administrations: a) determining whether the subject meets at least one criterion for shortening or lengthening one or more intervals between administrations of a VEGF receptor fusion protein by 2 weeks, 3 weeks, 4 weeks, or 2-4 weeks; b) if such a determination is made, administering additional doses of the VEGF receptor fusion protein with shortened or extended intervals between doses; The criteria for extending the interval are: 1. Loss of <5 letters of BCVA, and / or 2. The CRT is <300 or 320 micrometers, The criteria for shortening the interval are: 1. Loss of >10 letters of BCVA, 2. Persistence or worsening of DME, and / or 3. A VEGF receptor fusion protein is provided, which comprises an increase in CRT of >50 micrometers.

[0060] The present invention provides a VEGF receptor fusion protein for use in treating or preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof who has been previously treated with one or more 2 mg doses of the VEGF receptor fusion protein, comprising administering to the subject's eye a single initial administration of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration.

[0061] The present invention provides a VEGF receptor fusion protein for use in treating and preventing an intraocular neovascular disease in a subject in need thereof, comprising administering to the subject's eye at least 8 mg of the VEGF receptor fusion protein in one or more doses about every 24 weeks.

[0062] A VEGF receptor fusion protein for use in treating and preventing intraocular neovascular diseases, the treatment or prevention being packaged together prior to each administration. one single-dose glass vial with a protective plastic cap and stopper containing approximately 70 microliters of an aqueous formulation comprising 8 mg of a VEGF receptor fusion protein; one 18 gauge x 1.5 inch, 5 micron filter needle including tip and bevel; Next, (1) visually inspecting the aqueous formulation in the vial, and if particles, turbidity, or discoloration are visible, then using another vial of aqueous formulation containing a VEGF receptor fusion protein; (2) removing the protective plastic cap from the vial; (3) Clean the top of the vial with an alcohol wipe, and then Aseptic technique (4) removing an 18 gauge x 1.5 inch, 5 micron filter needle and a 1 mL syringe from their packaging; (5) attaching the filter needle to the syringe by twisting the filter needle onto the tip of the luer lock syringe; (6) pushing the filter needle into the center of the vial stopper until the needle is fully inserted into the vial and the tip of the needle contacts the bottom or bottom edge of the vial; (7) Withdrawing the entire contents of the VEGF receptor fusion protein vial into the syringe, holding the vial upright and slightly tilted while ensuring the bevel of the filter needle is submerged in the liquid; (8) tilting the vial during withdrawal to keep the bevel of the filter needle submerged in the formulation; (9) fully withdrawing the plunger rod when emptying the vial to completely empty the filter needle; (10) removing the filter needle from the syringe and discarding the filter needle; (11) removing a 30 gauge x 0.5 inch needle from its packaging and attaching the needle to the syringe by tightly twisting the needle onto the luer lock syringe tip; (12) Holding the syringe with the needle pointing upward, check for air bubbles in the syringe, and if there are any, gently flick the syringe with your finger until the bubbles rise to the top; (13) Slowly pushing the plunger out so that the plunger tip is aligned with the graduation mark marking the 70 microliter mark on the syringe.

[0063] The present invention provides a VEGF-receptor fusion protein for use in treating or preventing intraocular neovascular diseases, neovascularized age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein the subject is receiving a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema comprising a single initial dose of about 2 mg of the VEGF-receptor fusion protein, followed by four secondary doses of about 2 mg of the VEGF-receptor fusion protein, followed by one or more tertiary doses of about 2 mg of the VEGF-receptor fusion protein, each secondary dose administered about four weeks after the immediately preceding dose, and each tertiary dose administered about eight weeks after the immediately preceding dose, and the subject is at any stage (initial, secondary, or tertiary) of the 2 mg VEGF-receptor fusion protein dosing regimen.

[0064] The present invention provides a VEGF-receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF-receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising approximately 103 to 126 mg / mL of the VEGF-receptor fusion protein, a histidine-based buffer, and arginine.

[0065] The present invention provides a VEGF-receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF-receptor fusion protein is an aqueous pharmaceutical formulation comprising approximately 114.3 mg / ml of the VEGF-receptor fusion protein, a histidine-based buffer, and arginine.

[0066] The present invention provides aflibercept for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein ≥ 8 mg of aflibercept is present in the aqueous pharmaceutical formulation, and the aflibercept has less than about 3.5% high molecular weight species immediately after production and purification, and / or about 6% or less high molecular weight species after storage at about 2 to 8°C for about 24 months.

[0067] The present invention provides a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein ≥ 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation, the aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein; Approximately 10 to 100 mM L-arginine, Sucrose and a histidine-based buffer; a surfactant, The present invention provides a VEGF receptor fusion protein, wherein the formulation has a pH of about 5.0 to about 6.8, and the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification, and / or has about 6% or less high molecular weight species after storage at about 2 to 8°C for about 24 months.

[0068] The present invention relates to a VEGF receptor fusion protein for use in treating and preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein ≥ 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation, the aqueous pharmaceutical formulation comprising: Contains >100 mg / ml VEGF receptor fusion protein, histidine-based buffer, and L-arginine; 140 mg / ml aflibercept, 20 mM histidine-based buffer, 5% sucrose, 0.03% polysorbate 20, 10 mM L-arginine, pH 5.8, or 150±15 mg / ml aflibercept, 10 mM phosphate buffer, 8±0.8% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.9-6.5, or aflibercept at 103-126 mg / ml, 10 ± 1 mM histidine-based buffer, 5 ± 0.5% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20, and 50 ± 5 mM L-arginine, pH 5.5-6.1; 140 mg / ml aflibercept, 10 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8, or 114.3 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8, or Contains >100 mg / ml aflibercept, a histidine-based buffer, and L-arginine, or aflibercept at >100 mg / ml, with a pH of about 5.8, wherein the formulation forms <3% HMW aggregates after 2 months of incubation at 5°C; Approximately 114.3 mg / mL aflibercept, 10 mM to 50 mM histidine-based buffer, sugar, non-ionic surfactant, L-arginine, pH 5.8, or approximately 114.3 mg / mL aflibercept; 10 mM His / His-HCl buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, pH 5.8; or A VEGF receptor fusion protein containing approximately 114.3 mg / mL of aflibercept, arginine monohydrochloride, histidine, histidine hydrochloride monohydrate, polysorbate 20, sucrose, and water for injection is provided.

[0069] The present invention provides a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, the subject comprising: an improvement in the Diabetic Retinopathy Severity Scale (DRSS), e.g., by at least 2 or 3 stages; -Improvement of best corrected visual acuity, Dry retina, · Obtaining best corrected visual acuity, Acquisition of best-corrected visual acuity of at least 5, 10, or 15 letters; ·BCVA of at least 69 letters, ·Decreased central retinal thickness (CRT), Absence of vascular leakage as measured by fluorescein angiography (FA), Improvement from pre-treatment baseline in the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) total score, The retina is free of fluid (total fluid, intraretinal fluid [IRF] and / or subretinal fluid [SRF]) in the fovea and central subfields; Preservation of fluid-free retina (total fluid in the fovea and central subfields, IRF and / or SRF) Macular edema loss, Fluid-free retina on spectral-domain optical coherence tomography (SD-OCT), and / or and once initiated, no deviation from HDq12 or HDq16 treatment regimens is achieved and / or maintained.

[0070] The present invention provides a VEGF receptor fusion protein for use in treating and preventing intraocular neovascular disease, nAMD, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, the subject comprising: BVCA that is non-inferior to aflibercept administered intravitreally at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, then 2 mg approximately every 8 weeks or once every 2 months. A BCVA (according to ETDRS letter score) increase of approximately 7, 8, or 9 letters from the start of treatment to 60 weeks, with a baseline BCVA of approximately 61, 62, or 63. BCVA (according to ETDRS letter score) of at least approximately 69 letters by 48 or 60 weeks after the start of treatment; - No loss of BCVA of 5, 10, 15, or 69 letters or more at 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment. Improvement in BCVA (according to ETDRS letter score) by 12, 24, 36, 49, 60, 72, 84, or 90 weeks after initiation of treatment. -Improvement of BVCA by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after initiation of treatment. Between 48 and 60 weeks, the BCVA score (according to the ETDRS letter score) is approximately 69, 70, 71, 72, or 73. Between 36 and 48 weeks, a change in BCVA score (according to ETDRS letter score) from the start of treatment of approximately 7, 8, or 9, with a BCVA of approximately 60 or 70 at any time between 36 and 48 weeks. A change in BCVA score (according to ETDRS letter score) from the start of treatment of approximately 7, 8, or 9 between 48 and 60 weeks, with a BCVA of approximately 69, 70, 71, 72, or 73 at any time between 48 and 60 weeks; An increase in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or equivalent Snellen visual acuity of ≥ 4 letters, ≥ 5 letters, ≥ 6 letters, ≥ 7 letters, ≥ 8 letters, > 9 letters, or > 10 letters by week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 of treatment; - No loss of 5, 10, or 15 letters (according to ETDRS letter score) by 48 or 60 weeks of treatment Gain at least 5, 10, or 15 letters (according to ETDRS letter score) by week 48 or 60 of treatment. By 4 weeks after starting treatment, patients will have an improvement in BCVA of approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 8 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 12 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 16 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 20 weeks after starting treatment, BCVA will improve by approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or by approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 24 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 28 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 32 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 36 weeks after starting treatment, patients will have an improvement in BCVA of 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 40 weeks after starting treatment, patients will have an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 44 weeks after starting treatment, patients will have an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 48 weeks after starting treatment, patients will have an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. -Improvement in BCVA by approximately 8 weeks after the start of treatment, which is maintained throughout the treatment regimen until at least 48 weeks. By 4 weeks after starting treatment, the BCVA should be approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 66 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 8 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 67 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 12 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 16 weeks after initiation of treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 20 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 24 weeks after starting treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 67 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 28 weeks after initiation of treatment, the BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 32 weeks after initiation of treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 36 weeks after initiation of treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 40 weeks after starting treatment, the BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 44 weeks after initiation of treatment, BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By 48 weeks after initiation of treatment, the BCVA should be approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 regimen. By week 48 after treatment initiation, patients receiving an HDq12 regimen with a baseline BCVA of approximately <73 ETDRS letters will achieve an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity), By week 48 after treatment initiation, an improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq12 regimen and baseline BCVA is approximately >73 ETDRS letters, By week 48 after treatment initiation, an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq16 regimen and baseline BCVA is approximately <73 ETDRS letters, By week 48 after treatment initiation, an improvement in BCVA of approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq16 regimen and baseline BCVA is approximately >73 ETDRS letters, By week 48 after treatment initiation, an improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq12 regimen and baseline CRT is approximately <400 micrometers, By week 48 after treatment initiation, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq12 regimen and with a baseline CRT of approximately >400 micrometers, By week 48 after initiation of treatment, an improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) when receiving an HDq16 regimen and baseline CRT is approximately <400 micrometers, An improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) by 48 weeks after treatment initiation when receiving an HDq16 regimen and baseline CRT is approximately >400 micrometers. Achieving >5, >10, or ≥15 letters of BCVA (according to ETDRS letter score) by Week 12, 24, 36, 49, 60, 72, 84, or 90 of treatment; - An improvement of ≥2 or >3 steps in the Diabetic Retinopathy Severity Scale (DRSS) by 12, 24, 36, 49, 60, 72, 84, or 90 weeks after initiation of treatment; A ≥ 2-grade improvement in the Diabetic Retinopathy Severity Scale (DRSS) by Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 of treatment; - The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the fovea and central subfield by 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment, as measured by optical coherence tomography (OCT); - No vascular leakage as measured by fluorescein angiography (FA) at 12, 24, 36, 49, 60, 72, 84, or 90 weeks after initiation of treatment; Maintenance of a fluid-free retina (total fluid in the fovea and central subfield, IRF, and / or SRF) through 12, 24, 36, 49, 60, 72, 84, or 90 weeks from the start of treatment; A reduction in the total area of ​​fluorescein leakage within the ETDRS grid (mm²) of approximately 12, 13, or 14 mm² or more at 48 or 60 weeks, as measured by fluorescein angiography; - A fluid-free retina by spectral-domain optical coherence tomography (SD-OCT) at 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment; The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the fovea by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after the start of treatment; - Dry retina within 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment. The fovea is fluid-free by 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment, as measured by optical coherence tomography (OCT); A change in central retinal thickness of approximately -118 or -118.3 micrometers if receiving an HDq12 regimen, or approximately -124 or -125 or -124.9 or -125.5 micrometers if receiving an HDq16 regimen, by 4 weeks after starting treatment; A change in central retinal thickness of approximately -137 or -137.4 micrometers if receiving an HDq12 regimen, or approximately -139 or -140 or -139.6 or -140.3 micrometers if receiving an HDq16 regimen, by 8 weeks after starting treatment; A change in central retinal thickness of approximately 150 or -150.1 micrometers if receiving an HDq12 regimen, or approximately -152 or -153 or -152.7 or -153.4 micrometers if receiving an HDq16 regimen, by 12 weeks after starting treatment; A change in central retinal thickness of approximately 139 or -139.4 micrometers if receiving an HDq12 regimen, or approximately -145 or -146 or -145.5 or -146.4 micrometers if receiving an HDq16 regimen, by 16 weeks after starting treatment; A change in central retinal thickness of approximately -117 or -117.1 micrometers if receiving an HDq12 regimen, or approximately -112 or -113 or -112.5 or -113.3 micrometers if receiving an HDq16 regimen, by 20 weeks after starting treatment A change in central retinal thickness of approximately -158 or -158.1 micrometers if receiving an HDq12 regimen, or approximately -103 or -104 or -103.8 or -104.3 micrometers if receiving an HDq16 regimen, by 24 weeks after starting treatment; A change in central retinal thickness of approximately -146, -147, or -146.7 micrometers if receiving an HDq12 regimen, or approximately -162 or -162.3 micrometers if receiving an HDq16 regimen, by 28 weeks after starting treatment A change in central retinal thickness of approximately -132 micrometers if receiving an HDq12 regimen, or approximately -145, -146, or -145.8 micrometers if receiving an HDq16 regimen, by 32 weeks after starting treatment; A change in central retinal thickness of approximately -168 or -168.1 micrometers if receiving an HDq12 regimen, or approximately -124 or -125 or -124.7 or -125.2 micrometers if receiving an HDq16 regimen, by 36 weeks after starting treatment; A change in central retinal thickness of approximately -163 micrometers if receiving an HDq12 regimen, or approximately -122 or -123 or -122.5 or -123.1 micrometers if receiving an HDq16 regimen, by 40 weeks after starting treatment; A change in central retinal thickness of approximately -147, -148, or -147.4 micrometers if receiving an HDq12 regimen, or approximately -164, -164.1, or -164.3 micrometers if receiving an HDq16 regimen, by 44 weeks after starting treatment; A change in central retinal thickness of approximately -171, -172, or -171.7 micrometers if receiving an HDq12 regimen, or approximately -148, -149, -148.3, or -149.4 micrometers if receiving an HDq16 regimen, by 48 weeks after starting treatment; A change in central retinal thickness of approximately -181.95 or -176.24 micrometers if receiving an HDq12 regimen, or approximately -166.26 or -167.18 micrometers if receiving an HDq16 regimen, by 60 weeks after starting treatment; If receiving an HDq12 regimen, a change in central retinal thickness of approximately -118, -119, or -118.3 micrometers between the start of treatment (week 0) and week 4, A change in central retinal thickness of approximately -19, -20, or -19.1 micrometers between weeks 4 and 8 if receiving an HDq12 regimen, A change in central retinal thickness of approximately -12, -13, or -12.7 micrometers between weeks 8 and 12 if receiving an HDq12 regimen, A change in central retinal thickness of approximately -40 or -41 micrometers between weeks 20 and 24 when receiving the HDq12 regimen A change in central retinal thickness of approximately -36, -37, or -36.1 micrometers between weeks 32 and 36 if receiving an HDq12 regimen, A change in central retinal thickness of approximately -24, -25, or -24.3 micrometers between weeks 44 and 48 if receiving an HDq12 regimen, A change in central retinal thickness of -4, -5, or -4.5 micrometers between weeks 48 and 60 if receiving an HDq12 regimen, If receiving an HDq16 regimen, a change in central retinal thickness of approximately -124, -125, or -124.9 micrometers between the start of treatment (week 0) and week 4, A change in central retinal thickness of approximately -14, -15, or -14.7 micrometers between weeks 4 and 8 if receiving an HDq16 regimen, A change in central retinal thickness of approximately -13, -14, or -13.1 micrometers between weeks 8 and 12 if receiving an HDq16 regimen, A change in central retinal thickness of approximately -58, -59, or -58.5 micrometers between weeks 24 and 28 if receiving an HDq16 regimen, A change in central retinal thickness of approximately -41, -42, or -41.6 micrometers between weeks 40 and 44 if receiving an HDq16 regimen, a reduction in central retinal thickness by week 4, 5, 6, 7, or 8 after initiation of treatment that is maintained within approximately +17 micrometers, +18 micrometers, or +19 micrometers throughout the treatment regimen through at least week 48 of treatment; a reduction in central retinal thickness of approximately 100, 125, 150, 175, or 200 micrometers by 12, 24, 36, 49, 60, 72, 84, or 90 weeks after the start of treatment; A reduction in central retinal thickness of about 148 to 182 micrometers by about 48 or 60 weeks after the start of treatment, as measured by optical coherence tomography (OCT), with a baseline CRT of about 449, 450, 455, or 460 micrometers; a reduction in central retinal thickness (CRT) of at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 micrometers by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after the start of treatment; about 0.1667 days after the first dose, plasma free aflibercept is about 0.149 (±0.249) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.3333 days after the first dose, plasma free aflibercept is about 0.205 (±0.250) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 1 day after the first dose, plasma free aflibercept is about 0.266 (±0.211) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 2 days after the first dose, plasma free aflibercept is about 0.218 (±0.145) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 4 days after the first dose, plasma free aflibercept is about 0.140 (±0.0741) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 7 days after the first dose, plasma free aflibercept is about 0.0767 (±0.0436) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 14 days after the first dose, plasma free aflibercept is about 0.0309 (±0.0241) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 21 days after the first dose, plasma free aflibercept is about 0.0171 (±0.0171) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 28 days after the first dose, plasma free aflibercept is about 0.00730 (±0.0113) mg / L, where free aflibercept is undetectable in plasma at baseline and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.1667 days after the first dose, the plasma adjusted bound aflibercept is about 0.00698 (±0.0276) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.3333 days after the first dose, the plasma adjusted bound aflibercept is about 0.00731 (±0.0279) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 1 day after the first dose, the plasma adjusted bound aflibercept is about 0.0678 (±0.0486) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 2 days after the first dose, the plasma adjusted bound aflibercept is about 0.138 (±0.0618) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 4 days after the first dose, the plasma adjusted bound aflibercept is about 0.259 (±0.126) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 7 days after the first dose, the plasma adjusted bound aflibercept is about 0.346 (±0.151) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 14 days after the first dose, the plasma adjusted bound aflibercept is about 0.374 (±0.110) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 21 days after the first dose, the plasma adjusted bound aflibercept is about 0.343 (±0.128) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 28 days after the first dose, the plasma adjusted bound aflibercept is about 0.269 (±0.149) mg / L, and the baseline adjusted bound aflibercept is about 0.00583 mg / L (±0.0280), and the subject has not received intravitreal aflibercept treatment for at least 12 weeks; - the peak plasma concentration of free aflibercept is reached approximately 0.965 days after the first dose; - Reaching a peak plasma concentration of free aflibercept of approximately 0.310 mg / l (±0.263); -Free aflibercept in plasma is approximately 0 to approximately 1.08 mg / L. The maximum plasma free aflibercept concentration (mg / L) per mg aflibercept dose (mg) is approximately 0.388 (±0.0328) mg / L / mg. - Peak plasma concentrations of adjusted conjugated aflibercept are reached approximately 14 days after the first dose; Achieving a peak plasma concentration of approximately 0.387 mg / l (±0.135) of adjusted bound aflibercept; - adjusted bound aflibercept in plasma is approximately 0.137 to approximately 0.774 mg / L; The maximum plasma adjusted bound aflibercept (mg / l) per mg aflibercept dose is approximately 0.483 (±0.0168) mg / l / mg; - Absence of anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment Improvement from pre-treatment baseline in the National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score, and / or Achieving and / or maintaining one or more of the following: loss of macular edema.

[0071] The present invention provides a VEGF-receptor fusion protein for use in treating and preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein the interval between doses of 8 mg of the VEGF-receptor fusion protein is adjusted (increased / maintained / shortened) based on visual and / or anatomical outcomes.

[0072] The present invention provides VEGF-receptor fusion proteins for use in treating and preventing neovascular age-related macular degeneration, diabetic retinopathy, and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF-receptor fusion protein is administered according to an as-needed (PRN (pro re nata)), capped PRN, or Treat and Extend (T&E) dosing regimen, as needed.

[0073] The present invention also provides a kit comprising: i) a container containing a VEGF receptor fusion protein, preferably aflibercept; and ii) instructions for use of the VEGF fusion protein. In one embodiment of the present invention, the container is a vial or a prefilled syringe. The vial is a type I glass vial containing a nominal fill volume of about 0.26 mL of solution for intravitreal injection. In one embodiment of the present invention, the container contains a VEGF receptor fusion protein at a concentration of 100 mg / mL or more, or the container contains aflibercept at a concentration of about 114.3 mg / mL. In one embodiment of the present invention, the instructions for use include instructions for use of the VEGF fusion protein or aflibercept for the treatment of DME and / or AMD. In one embodiment of the present invention, the instructions for use indicate that: i) the container contains 8 mg (114.3 mg / mL) aflibercept solution for intravitreal injection; ii) each single-dose vial provides a quantity usable to deliver a single 70 microliter dose containing 8 mg aflibercept to an adult patient; iii) the recommended dose is 8 mg aflibercept (equivalent to 70 microliters of injection solution); iv) the 8 mg aflibercept treatment is administered as three consecutive doses; v) the injection interval may then be extended up to every 16 or 20 weeks; vi) the treatment interval may be adjusted based on the physician's visual and / or anatomical outcome judgment; and / or vii) 8 mg aflibercept / 0.07 mL is provided as a sterile, aqueous solution containing arginine monohydrochloride, histidine, histidine hydrochloride monohydrate, polysorbate 20, sucrose, and water for injection. [Brief explanation of the drawings]

[0074] [Figure 1] Figure 1 provides an overview of the PHOTON clinical trial. [Figure 2] Figure 2 shows the key eligibility criteria (inclusion and exclusion criteria) for the PHOTON clinical trial. [Figure 3] Figure 3 shows the administration schedule and dose regimen change (DRM) criteria for the PHOTON clinical trial (up to week 48). [Figure 4] Figure 4 shows the criteria for changing the administration regimen in the PHOTON clinical trial. [Figure 5] Figure 5 shows patient disposition at week 48 in the PHOTON clinical trial. [Figure 6] FIG. 6 shows the baseline demographic data of subjects in the PHOTON clinical trial. [Figure 7] Figure 7 shows the baseline characteristics of the study eye of the subjects in the PHOTON clinical trial. [Figure 8] Figure 8 shows the mean number of injections up to week 48 in the PHOTON clinical trial. [Figure 9] Figure 9 shows the mean change in best corrected visual acuity (BCVA) from baseline to week 48 in the PHOTON clinical trial. The least squares mean change from baseline to week 48 is shown. [Figure 10] FIG. 10 shows the percentage of subjects maintaining Q12-week and Q16-week intervals through week 48 in the PHOTON clinical trial. [Figure 11] FIG. 11 is the key secondary endpoint (EP), proportion of subjects with a ≧2-grade improvement in the Diabetic Retinopathy Severity Scale (DRSS) at Week 48 in the PHOTON clinical trial. [Figure 12] FIG. 12 shows the percentage of subjects without foveal retinal fluid at 48 weeks in the PHOTON clinical trial. [Figure 13] Figure 13 shows the mean change from baseline in central retinal thickness to week 48 in the PHOTON clinical trial. Several consistent intervals are highlighted in the three insets. [Figure 14A] Figures 14A, 14B, and 14C show the ocular serious treatment-emergent adverse events (TEAEs) through 48 weeks (Figure 14A), the most common adverse events (AEs) through 48 weeks (Figure 14B), and non-ocular safety through 48 weeks (Figure 14C) in the PHOTON clinical trial. [Figure 14B] Same as above. [Figure 14C] Same as above. [Figure 15]FIG. 15 shows treatment-emergent intraocular inflammation through 48 weeks in the PHOTON clinical trial. [Figure 16] FIG. 16 shows the mean change from baseline in intraocular pressure through week 48 in the PHOTON clinical trial. [Figure 17] FIG. 17 shows the percentage of subjects meeting intraocular pressure criteria in the PHOTON clinical trial. [Figure 18] Figure 18: ≧1% non-ocular serious TEAEs through Week 48 in the PHOTON clinical trial. [Figure 19] FIG. 19 shows the Anti-Platelet Trialists' Collaboration (APTC)-defined events that occurred treatment-emergently through week 48 in the PHOTON clinical trial. [Figure 20] FIG. 20 shows treatment-emergent hypertensive events through 48 weeks in the PHOTON clinical trial. [Figure 21] FIG. 21 shows the potentially clinically significant values ​​(PCVs) of blood pressure up to week 48 in the PHOTON clinical trial. [Figure 22] Figure 22 shows the mean change from baseline in systolic blood pressure through week 48 in the PHOTON clinical trial. SBP from baseline to week 9 and mean baseline SBP are shown in the inset table. [Figure 23] Figure 23 shows the mean change from baseline in diastolic blood pressure in the PHOTON clinical trial through week 48. DBP from baseline to week 9 and mean baseline DBP are shown in the inset table. [Figure 24] Figure 24 shows mortality up to 48 weeks in the PHOTON trial. [Figure 25A]Figures 25A and 25B show (A) the mean change from baseline in BCVA score (ETDRS letter) in the study eye to Week 60 in the OC (full analysis set). (B) the least squares mean change from baseline in BCVA score (ETDRS letter) in the study eye to Week 60 (full analysis set). Abbreviations: 2q8: aflibercept 2 mg administered every 8 weeks after five initial injections at 4-week intervals; HDq12: high-dose aflibercept 8 mg administered every 12 weeks after three initial injections at 4-week intervals; HDq16: high-dose aflibercept 8 mg administered every 16 weeks after three initial injections at 4-week intervals. BCVA = best-corrected visual acuity, ETDRS = Early Treatment Diabetic Retinopathy Study, HD = high dose, OC = observed cases, SE = standard error. OC: Observations after the ICE specified for the primary estimand were excluded. [Figure 25B] Same as above. [Figure 26A] Figures 26A and 26B show (A) the mean change from baseline in central retinal thickness (microns) by visit to Week 60 in the OC (full analysis set). (B) the least squares mean change from baseline in central retinal thickness (microns) by visit to Week 60 in the OC (full analysis set). Abbreviations: 2q8 = aflibercept 2 mg administered every 8 weeks after five initial injections at 4-week intervals; HDq12 = high-dose aflibercept 8 mg administered every 12 weeks after three initial injections at 4-week intervals; HDq16 = high-dose aflibercept 8 mg administered every 16 weeks after three initial injections at 4-week intervals. ICE = intercurrent event; OC = observed case; SE = standard error. OC = observations after the ICE specified for the primary estimand were excluded. [Figure 26B] Same as above. [Figure 27]Figure 27 shows the mean (+SD) plasma free aflibercept concentrations (mg / L) by nominal time and treatment in subjects with DME who received unilateral treatment in the Dense PK Sampling substudy (VGFTe-HD-DME-1934 study, log scale, [DPKS]). N = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = aflibercept 2 mg administered intravitreally every 8 weeks after five monthly initial doses; HDq12 = high-dose (8 mg) aflibercept administered intravitreally every 12 weeks after three monthly initial doses; HDq16 = high-dose (8 mg) aflibercept administered intravitreally every 16 weeks after three monthly initial doses; LLOQ = lower limit of quantification; D = day; H = hour. Note: The table below the figure shows the number of subjects contributing to the statistics for the corresponding visit / timepoint and treatment. Concentrations below the LLOQ were set at LLOQ / 2. HDq12+HDq16=combined data from the HDq12 and HDq16 treatment groups. Patients in the dense PK sub-study received aflibercept injection in only one eye. [Figure 28]Figure 28 shows the mean (+SD) adjusted plasma concentration of bound aflibercept (mg / L) by nominal time and treatment in subjects with DME who received unilateral treatment in the Dense PK Sampling substudy (VGFTe-HD-DME-1934 study, log scale, [DPKS]). N = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = aflibercept 2 mg administered intravitreally every 8 weeks after five monthly initial doses; HDq12 = high-dose (8 mg) aflibercept administered intravitreally every 12 weeks after three monthly initial doses; HDq16 = high-dose (8 mg) aflibercept administered intravitreally every 16 weeks after three monthly initial doses; LLOQ = lower limit of quantification; D = day; H = hour. Note: The table below the figure shows the number of subjects contributing to the statistics for the corresponding visit / timepoint and treatment. Concentrations below the LLOQ were set at LLOQ / 2. Adjusted bound aflibercept = 0.717 * bound aflibercept. HDq12 + HDq16 = combined data from the HDq12 and HDq16 treatment arms. Patients in the dense PK sub-study received aflibercept injection in only one eye. [Figure 29]Figure 29 shows the mean (+SD) plasma free aflibercept concentrations (mg / L) by nominal time and treatment group in subjects with DME in the Sparse PK Sampling Study (VGFTe-HD-DME-1934, log scale, [PKAS]). N = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = 2 mg aflibercept administered intravitreally every 8 weeks after five monthly initial doses; HDq12 = high-dose (8 mg) aflibercept administered intravitreally every 12 weeks after three monthly initial doses; HDq16 = high-dose (8 mg) aflibercept administered intravitreally every 16 weeks after three monthly initial doses; LLOQ = lower limit of quantification; PKAS = pharmacokinetic analysis set. Note: The table below the figure shows the number of subjects contributing to the statistics for the corresponding visit / timepoint and treatment group. Concentrations below the LLOQ were set at LLOQ / 2. Post-dose samples and samples collected during the multi-point PK substudy (post-dose on Day 0 through Day 21) were excluded. [Figure 30]Figure 30 shows the mean (+SD) adjusted bound aflibercept concentrations (mg / L) in plasma by nominal time and treatment group in subjects with DME in the Sparse PK Sampling Study (VGFTe-HD-DME-1934, log scale, [PKAS]). N = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = 2 mg aflibercept intravitreally every 8 weeks after five initial doses monthly; HDq12 = high-dose (8 mg) aflibercept intravitreally every 12 weeks after three initial doses monthly; HDq16 = high-dose (8 mg) aflibercept intravitreally every 16 weeks after three initial doses monthly; LLOQ = 3 / 4 lower limit of quantitation. Note: The table below the figure shows the number of subjects contributing to the statistics for the corresponding visit / timepoint and treatment group. Concentrations below the LLOQ were set at LLOQ / 2. Adjusted bound aflibercept = 0.717 * bound aflibercept. Post-dose samples and samples collected during the dense PK sub-study (post-dose on Day 0 through Day 21) were excluded. [Figure 31] Figure 31 shows a structural representation of the population pharmacokinetic model for aflibercept after IV, SC, and IVT administration. CMT = compartment, IV = intravenous, IVT = intravitreal, K = elimination rate constant for free aflibercept, K = elimination rate constant for adjusted bound aflibercept, K = absorption rate from the subcutaneous depot compartment, K = elimination rate constant from the tissue (platelet) compartment, Q = intercompartmental clearance of free aflibercept between the ocular and central compartments, QF = intercompartmental clearance of free aflibercept, VMK = saturable Michaelis-Menten binding of free aflibercept to VEGF, VMK = saturable elimination from the plasma compartment to the tissue compartment (platelets), and CMT = both represent the plasma compartment, assumed to be of equal volume. [Figure 32]FIG. 32. Mean (+SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days for single 2 mg and 8 mg IVT doses of aflibercept in nAMD or DME in the Dense PK Sub-study (DPKS, log scale). LQ = below limit of quantitation, DME = diabetic macular edema, DPKS = dense pharmacokinetic sub-study, HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections monthly, HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections monthly, IVT = intravitreal, LLOQ = lower limit of quantitation, N = number of subjects, nAMD = neovascular age-related macular degeneration, SD = standard deviation, adjusted bound aflibercept = 0.717 * bound aflibercept Note: Concentrations below the LLOQ (0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Note: The first 28 days of 8 mg HD aflibercept data (obtained from PULSAR or PHOTON) are composite data from subjects receiving HDq12 or HDq16. One PULSAR subject with an outlier free aflibercept concentration greater than 10-fold the mean concentration at day 28 is excluded. Records after fellow eye treatment are excluded. Data source: Drug concentration data from the 48-week database lock for PULSAR and PHOTON, and the final lock for CANDELA. [Figure 33]Figure 33 shows the observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in plasma over 28 days after a single IVT injection in subjects with nAMD or DME in the Dense PK Sub-study (DPKS), stratified by dose and population. DME = diabetic macular edema, IVT = intravitreal, LLOQ = lower limit of quantification, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetic. The observed concentrations below the lower limit of quantification (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set at LLOQ / 2. Data source: Drug concentration data from the dense PK sub-study in PHOTON, PULSAR, and CANDELA. [Figure 34] Figure 34 is an overlay of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in plasma for a combined population of nAMD and DME. 2q8 = aflibercept 2 mg administered every 8 weeks after three initial injections spaced 4 weeks apart; 2q12 = aflibercept 2 mg administered every 12 weeks after three initial injections spaced 4 weeks apart; DME = diabetic macular edema; HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections spaced 4 weeks apart; HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections spaced 4 weeks apart; IVT = intravitreal; LLOQ = lower limit of quantification; nAMD = neovascular age-related macular degeneration. The observed concentrations below the lower limit of quantitation (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Data source: Drug concentration data from CANDELA, PHOTON, and PULSAR. [Figure 35]35 shows the model-predicted exposure (mg) of aflibercept after a single IVT injection stratified by dosing regimen in subjects with a combined nAMD and DME. DME = diabetic macular edema, HD = aflibercept 8 mg, IVT = intravitreal, nAMD = neovascular age-related macular degeneration, PI = prediction interval, PK = pharmacokinetics, QE = intercompartmental clearance of free aflibercept between the ocular compartment and the central compartment. The adjusted LLOQ (0.0624 μg) is set as the LLOQ of free aflibercept in plasma (i.e., 0.0156 mg / L) × the assumed volume of the test ocular compartment (i.e., 4 mL) in the PK model. In the clinical trials included in the population PK analysis dataset, aflibercept concentrations (free or bound) were not measured in the study eye, so this target was arbitrarily selected based on the LLOQ in plasma and used as a reference for comparison between dosing regimens to evaluate the effect of HD aflibercept on QE. [Figure 36]FIG. 36. Mean (+SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days, no outliers, for single 2 mg and 8 mg IVT doses of aflibercept in subjects with nAMD in the Dense PK Sub-study (DPKS, log scale). DME = diabetic macular edema, DPKS = dense pharmacokinetic sub-study, HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections monthly, HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections monthly, IVT = intravitreal, LLOQ = lower limit of quantification, N = number of subjects, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetic, SD = standard deviation, adjusted bound aflibercept = 0.717 * bound aflibercept. Note: Concentrations below the LLOQ (0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Note: The first 28 days of 8 mg data (obtained from PULSAR or PHOTON) are composite data from subjects receiving HDq12 or HDq16. Data source: Drug concentrations from the 48-week lock for PULSAR and PHOTON, and the final lock for CANDELA. Records after fellow eye treatment are excluded. [Figure 37]Figure 37. Mean (+SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days for single 2 mg and 8 mg IVT doses of aflibercept in subjects with nAMD in the Dense PK Sub-study (DPKS, log scale), with no outliers. DME = diabetic macular edema, DPKS = dense pharmacokinetic sub-study, HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections monthly, HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections monthly, IVT = intravitreal, LLOQ = lower limit of quantification, N = number of subjects, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetic, SD = standard deviation, adjusted bound aflibercept = 0.717 * bound aflibercept Note: Concentrations below the lower limit of quantification (LLOQ, 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set at LLOQ / 2. Data sources: Drug concentrations from the 48-week lock for PULSAR and the final lock for CANDELA. Data from VGFTOD-0702 will be included as reference (PK substudy concentrations will be subtracted by pre-dose concentrations if >LLOQ). Records after fellow eye treatment will be excluded. [Figure 38]FIG. 38. Mean (+SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days for single 2 mg and 8 mg IVT doses of aflibercept in subjects with DME in the Dense PK Sub-study (DPKS, log scale). BLQ = below limit of quantitation, DME = diabetic macular edema, DPKS = dense pharmacokinetic population, HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections monthly, HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections monthly, IVT = intravitreal, LLOQ = lower limit of quantitation, N = number of subjects, nAMD = neovascular age-related macular degeneration, SD = standard deviation. Note: Concentrations below the LLOQ (0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Adjusted bound aflibercept = 0.717 * bound aflibercept. Note: The first 28 days of 8mg data (obtained from PULSAR or PHOTON) are composite data from subjects receiving HDq12 or HDq16. Note: If a subject took aflibercept within 12 weeks before starting study drug and the baseline concentration is >BLQ, the concentration will be subtracted by the baseline concentration. Data source: Drug concentration data from week 48 lock in PHOTON. Drug concentration data (historical data) from VGFT-OD-0706 is included as a reference. Records after fellow eye treatment are excluded. [Figure 39]Figure 39 is an overlay of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in plasma for subjects with nAMD. 2q8 = aflibercept 2 mg administered every 8 weeks after three initial injections spaced 4 weeks apart; 2q12 = aflibercept 2 mg administered every 12 weeks after three initial injections spaced 4 weeks apart; DME = diabetic macular edema; HDq12 = aflibercept 8 mg administered every 12 weeks after three initial injections spaced 4 weeks apart; HDq16 = aflibercept 8 mg administered every 16 weeks after three initial injections spaced 4 weeks apart; IVT = intravitreal; LLOQ = lower limit of quantification; nAMD = neovascular age-related macular degeneration; PK = pharmacokinetic. Observed concentrations below the lower limit of quantitation (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set at LLOQ / 2. Data from PULSAR and CANDELA. [Figure 40] Figure 40 is an overlay of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in plasma for subjects with diabetic macular edema in the PHOTON study. 2q8 = aflibercept 2 mg administered every 8 weeks after three initial injections spaced 4 weeks apart, HDql2 = aflibercept 8 mg administered every 12 weeks after three initial injections spaced 4 weeks apart, HDql6 = aflibercept 8 mg administered every 16 weeks after three initial injections spaced 4 weeks apart, IVT = intravitreal, LLOQ = lower limit of quantification, data from PHOTON. [Figure 41] Figure 41 shows the dosing schedule and dosing regimen change (DRM) criteria for the PULSAR clinical trial (up to week 48). [Figure 42] Figure 42 shows the key eligibility criteria (inclusion and exclusion criteria) for the PULSAR clinical trial. [Figure 43] Figure 43 shows the dosing schedule and dosing regimen modification (DRM) criteria for the PULSAR clinical trial. [Figure 44] Figure 44 shows the dosing regimen modification (DRM) criteria for the PULSAR clinical trial. [Figure 45] Figure 45 shows patient disposition at week 48 in the PULSAR clinical trial. [Figure 46] Figure 46 shows the baseline demographic data of subjects in the PULSAR clinical trial. [Figure 47] FIG. 47 shows the baseline characteristics of the study eye of subjects in the PULSAR clinical trial. [Figure 48] Figure 48 shows the mean number of injections through week 48 in the PULSAR clinical trial. [Figure 49] Figure 49 shows the mean change in best corrected visual acuity (BCVA) in the PULSAR clinical trial through week 48. The least squares mean change from baseline at week 48 is shown in the table. [Figure 50] Figure 50 shows the percentage of subjects maintaining Q12 week and Q16 week intervals through week 48 in the PULSAR clinical trial. [Figure 51] Figure 51 is the key secondary endpoint, percentage of subjects without retinal fluid in the central subfield at week 16 in the PULSAR clinical trial. [Figure 52] Figure 52 is the percentage of subjects without retinal fluid in the central subfield at week 48 in the PULSAR clinical trial. [Figure 53A] Figure 53 (A-B) shows the mean change in central retinal thickness from baseline to week 48 (A), and central retinal thickness to week 48 (B). [Figure 53B] Same as above. [Figure 54A] Figure 54 (A-B) shows the serious ocular treatment-emergent adverse events (TEAEs) through Week 48 (A) and the most common adverse events (AEs) through Week 48 (B) in the PULSAR clinical trial. [Figure 54B] Same as above. [Figure 55] Figure 55 shows treatment-emergent intraocular inflammation through 48 weeks in the PULSAR clinical trial. [Figure 56]Figure 56 shows the mean change from baseline in intraocular pressure through week 48 in the PULSAR clinical trial. [Figure 57] Figure 57 shows the percentage of subjects meeting intraocular pressure criteria in the PULSAR clinical trial. [Figure 58] Figure 58 is non-ocular serious TEAEs > 0.5% through Week 48 in the PULSAR clinical trial. [Figure 59A] Figure 59 (A-B) shows the Anti-Platelet Trialists' Collaboration (APTC)-defined events that emerged from treatment up to week 48 in the PULSAR clinical trial (A) and non-ocular safety up to week 48 (B). [Figure 59B] Same as above. [Figure 60] Figure 60 shows treatment-emergent hypertensive events through 48 weeks in the PULSAR clinical trial. [Figure 61] Figure 61 shows the potentially clinically significant values ​​(PCVs) of blood pressure up to week 48 in the PULSAR clinical trial. [Figure 62] Figure 62 shows the mean change from baseline in systolic blood pressure through week 48 in the PULSAR clinical trial. The mean change from baseline to week 9 and the mean baseline blood pressure are shown in the inset table. [Figure 63] Figure 63 shows the mean change from baseline in diastolic blood pressure through week 48 in the PULSAR clinical trial. The mean change from baseline to week 9 and the mean baseline blood pressure are shown in the inset table. [Figure 64] Figure 64 shows mortality up to 48 weeks in the PULSAR clinical trial. [Figure 65] Figure 65 shows the PULSAR administration schedule up to week 60. The criteria for changing the administration regimen are listed in the inset. [Figure 66]Figure 66 shows absolute BCVA and change in BCVA from baseline (ETDRS letters) [PULSAR] to Week 60. Least squares mean change from baseline at Week 60 is shown. [Figure 67] Figure 67 shows the percentage of PULSAR patients who maintain HDq12 (8q12) and HDq16 (8q16) intervals through 60 weeks. [Figure 68] Figure 68 shows the mean number of injections [PULSAR] up to week 60 in each group. [Figure 69A] Figure 69 (A-C) shows central retinal thickness (CRT) and change from baseline [PULSAR] to week 60. (A) Central retinal thickness (micrometers) over time (observed values ​​- censored after ICE), (B) Mean change from baseline in CST (central subfield retinal thickness (interchangeable with CRT), micrometers) by visit to week 60, OC before ICE in the full analysis set, (C) LSmean (95% CI) change from baseline in CST (micrometers) by visit, MMRM (mixed model for repeated measures) in the full analysis set (through week 48). [Figure 69B] Same as above. [Figure 69C] Same as above. [Figure 70A] Figure 70 (A-G) summarizes the PULSAR safety data for PULSAR: (Figure 70A) ≥ 2% ocular TEAEs through Week 60, (Figure 70B) serious ocular TEAEs through Week 60, (Figure 70C) ≥ 2% non-ocular TEAEs through Week 60, (Figure 70D) ≥ 0.5% serious non-ocular TEAEs through Week 60, (Figure 70E) deaths through Week 60, (Figure 70F) mean change in systolic blood pressure at Week 60, and (Figure 70G) mean change in diastolic blood pressure at Week 60. [Figure 70B] Same as above. [Figure 70C] Same as above. [Figure 70D] Same as above. [Figure 70E] Same as above. [Figure 70F] Same as above. [Figure 70G] Same as above. [Figure 71] Figure 71 shows the 5, 10, and 15 letter changes at week 60. Observed (OC) (data censored after ICE) [PULSAR]. [Figure 72] Figure 72 is the % proportion of PULSAR subjects free of retinal fluid in the central subfield by visit (week and schedule of administration indicated) through Week 60. LOCF (data censored after ICE). [Figure 73] Figure 73 shows the percentage of PHOTON patients who maintained or extended their intervals through Week 96. Patients completed at Week 96. Values ​​may not add up to 100% due to rounding. Q8 is every 8 weeks, Q12 is every 12 weeks, Q16 is every 16 weeks, Q20 is every 20 weeks, and Q24 is every 24 weeks. [Figure 74A] Figure 74 (A & B) shows the BCVA and CRT over time for the patient in PULSAR Case Report 1. (A) Patient characteristics are shown along with images of the patient's retina at baseline, week 12, and week 96. A timeline of the absolute BCVA and absolute CRV achieved by the patient is provided at the bottom. The specific maintenance dosing interval (q16, q20, or q24) to which the patient was assigned at various time points is indicated. Dosing regimen change assessments were performed at the boxed weeks. (B) The interval to which the patient was assigned in the overall PULSAR subject population. [Figure 74B] Same as above. [Figure 75A] Figure 75 (A & B) shows the BCVA and CRT over time for the patient in PULSAR Case Report 2. (A) Patient characteristics are shown along with patient retinal images at baseline, week 12, and week 96. A timeline of the absolute BCVA and absolute CRV achieved by the patient is provided at the bottom. The specific maintenance dosing intervals (q12 and q16) to which the patient was assigned at various time points are indicated. Dosing regimen change assessments were performed at the boxed weeks. (B) Patients' assigned intervals for the entire PULSAR subject population. [Figure 75B] Same as above. [Figure 76] Figure 76 shows the BCVA and CRT over time for a patient from the PHOTON case report. (A) Patient characteristics are shown along with images of the patient's retina at baseline, week 12, and week 96. A timeline of the absolute BCVA and absolute CRV achieved by the patient is provided at the bottom. The specific maintenance dosing intervals (q16, q20, and q24) to which the patient was assigned at various time points are indicated. Dosing regimen change assessments were performed at the boxed weeks. DETAILED DESCRIPTION OF THE INVENTION

[0075] The present invention, in part, provides a safe and effective IVT injection of high-dose aflibercept that extends the maintenance dosing interval beyond 8 weeks and has at least similar functional and potentially improved anatomical outcomes. This regimen unexpectedly demonstrated a high degree of persistence in subjects, beyond what would be expected simply based on the higher doses of aflibercept administered.

[0076] EYLEA has become the standard of care for neovascular age-related macular degeneration (nAMD), diabetic macular edema (DME), and diabetic retinopathy (DR). EYLEA is prescribed for DME and DR at a monthly dose of 2 mg for five doses, followed by maintenance doses every eight weeks. The dosing regimen demonstrated a significantly higher percentage of subjects maintained on the 12- and 16-week dosing intervals. In clinical trials testing these dosing regimens, nearly 90% of subjects with diabetic macular edema were maintained on the 16-week dosing regimen. These durability data, coupled with a safety profile consistent with that of EYLEA, support high-dose aflibercept as a potential new standard of care for intraocular neovascular diseases such as DR or DME. The data presented herein demonstrated that 12- and 16-week dosing regimens of aflibercept 8 mg reached high barriers and sustained improvements in visual acuity and anatomical measures of retinal fluid over 48 weeks in subjects with diabetic macular edema. All of these results were achieved in subjects who were rapidly initiated on an extended dosing interval, the majority without the need for regimen changes. Overall, the pivotal study data support aflibercept 8 mg as offering a longer duration of action while maintaining a safety profile similar to that of EYLEA.

[0077] Prior to initiating the HD aflibercept clinical development program, pharmacokinetic simulations of the concentration-time profile of free aflibercept in the human vitreous using a one-compartment eye model predicted that IVT administration of 8 mg aflibercept could extend the dosing interval by approximately 20 days (two half-lives) compared with IVT administration of 2 mg. The HDq12 and HDq16 aflibercept regimens demonstrated longer durations of efficacy than predicted in HD clinical trials. A subsequent population PK analysis combining data from the CANDELA, PHOTON, and PULSAR Phase 3 trials predicted that the ocular clearance of free aflibercept was 34% slower for the HD aflibercept formulation compared with 2 mg aflibercept administered IVT as the EYLEA formulation, and predicted that the slower ocular clearance of HD aflibercept would result in both a longer duration of free aflibercept in the eye and an approximately 6-week longer duration of efficacy compared with the 2 mg formulation. The magnitude of the reduction in ocular clearance for the HD aflibercept drug compared to the 2 mg Eylea drug was greater than expected and was attributed to a statistically significant, high effect in this population PK model that could not be explained simply by the increase in dose from 2 mg to 8 mg, the "HD aflibercept drug effect."

[0078] Population PK estimated the median time to reach the lower limit of quantitation (LLOQ) for plasma free aflibercept concentrations after 2 mg IVT aflibercept to be 1.5 weeks compared with 3.50 weeks for 8 mg HD aflibercept. The longer period of systemic exposure to free aflibercept, representing movement of free aflibercept from the eye, with the HD aflibercept regimen is due not only to the higher dose administered and nonlinear systemic target-mediated elimination, but also to a 34% slower ocular clearance of free aflibercept. The slower ocular clearance of the HD aflibercept formulation was predicted to provide a 6-week longer duration of efficacy compared with the 2 mg aflibercept drug product, because the population PK estimated time to achieve free aflibercept levels in the ocular compartment for the 2q8 regimen at the end of the 8-week dosing interval occurs 6 weeks later with the HD aflibercept drug product. Exposure-response analysis estimated that the slower ocular clearance of 8 mg aflibercept due to the HD drug effect resulted in a dose regimen modification rate (DRM) that was 20.6% lower than what would be expected if the HD drug had the same ocular clearance as 2 mg aflibercept.

[0079] Standard methods in molecular biology are described by Sambrook, Fritsch and Maniatis (1982 & 1989 2 nd Edition, 2001 3 rd Edition)Molecular Cloning,A Laboratory Manual,Cold Spring Harbor Laboratory Press,Cold Spring Harbor,NY,Sambrook and Russell (2001)Molecular Cloning,3 rded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, Calif. Standard methods can also be found in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. New York, NY, which describes cloning and DNA mutagenesis in bacterial cells (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3), and bioinformatics (Vol. 4).

[0080] General methods for protein purification, including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization, have been described (Coligan et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and protein glycosylation are described (see, e.g., Coligan et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, Mo.; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, NJ, pp. 384-391). The production, purification, and fragmentation of polyclonal and monoclonal antibodies have been described (Coligan et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard techniques for characterizing ligand / receptor interactions are available (see, e.g., Coligan et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

[0081] Methods for flow cytometry, including fluorescence-activated cell sorting (FACS), are available (e.g., Owens et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2 nd (See, e.g., Molecular Probes (2003) Catalogue, Molecular Probes, Inc., Eugene, Oreg.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ.) Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, for use as diagnostic reagents, are available (Molecular Probes (2003) Catalogue, Molecular Probes, Inc., Eugene, Oreg.; Sigma-Aldrich (2003) Catalogue, St. Louis, Mo.).

[0082] Standard methods for immune system histology have been described (see, e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, Pa.; Louis et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0083] "Isolated" VEGF antagonists and VEGF receptor fusion proteins (e.g., aflibercept), polypeptides, polynucleotides, and vectors are at least partially free of other biological molecules from the cells or cell cultures in which they are produced. Such biological molecules include nucleic acids, proteins, other VEGF antagonists and VEGF receptor fusion proteins, lipids, carbohydrates, or other substances, such as cell debris and growth medium. Isolated VEGF antagonists or VEGF receptor fusion proteins may also be at least partially free of expression system components, such as biological molecules of host cells or their growth medium. In general, the term "isolated" is not intended to refer to the complete absence of such biological molecules (though small or trace amounts of impurities may remain), or the absence of water, buffers, or salts, or components of pharmaceutical formulations containing the VEGF antagonists or VEGF receptor fusion proteins.

[0084] Subject and patient are used interchangeably herein. The subject or patient is a mammal, such as a human, a mouse, a rabbit, a monkey, or a non-human primate, preferably a human. The subject or patient is said to be "suffering" from an intraocular neovascular disease, such as nAMD. Such a subject or patient has damage to one or both eyes. In an embodiment of the present invention, the subject or patient (preferably a human) has the following characteristics (currently or previously): 1. Age ≥ 50 years, e.g., 61, 62, 63, 74, or 75 years; 2. Have active subfoveal CNV secondary to nAMD, including parafoveal lesions affecting the fovea of ​​the eye; 3. Early treatment of diabetic retinopathy with best corrected visual acuity (BCVA) (ETDRS) letter score of about 78-24, 73-78, <73, 58, 59, 60, 61, 62, or 63 (corresponding to Snellen visual acuity of 20 / 40, 20 / 63, 20 / 50, 20 / 32, or 20 / 320), e.g., due to DME or wet AMD; 4. DME with central involvement of the eye with a central retinal thickness of ≥ 300 micrometers or ≥ 320 micrometers, or about 367, 368, 369, 370, 450, 451, 452, 453, 454, or 455 micrometers, and / or a CRT of ≥ 300 micrometers (or ≥ 320 micrometers on Spectralis), 5. Total lesion area is approximately 6 or 7 mm 2 For example, the lesion type is Occult type, predominantly classic type, or minimally classic type; 6. A DRSS score of level 43 or above or level 47 or below. 7. Type 1 diabetes or type 2 diabetes (insulin-dependent or non-insulin-dependent) (e.g., for more than about 15 years); 8. Hemoglobin A1C (%) of about 7 or 8 or more; 9. A body mass index of about 30 or 31 or greater; and / or 10. History of diabetic retinal edema, diabetic retinopathy, dry eye, vitreous detachment, retinopathy hypersensitivity, retinal hemorrhage, cataract surgery, retinal laser coagulation therapy, intraocular lens implant, or hypertension. and / or have one or more of the following characteristics: 1. Evidence of macular edema of any cause other than diabetes mellitus in the eye, 2. IOP ≥ 25mmHg in the eye; 3. The patient has had glaucoma filtration surgery in the past, or may need filtration surgery in the future. 4. Evidence of infectious blepharitis, keratitis, scleritis, or conjunctivitis in either eye within 4 weeks (28 days) of treatment, 5. Any intraocular inflammation and / or ocular infection in the eye within 12 weeks (84 days) of treatment, 6. History of idiopathic or autoimmune uveitis of the eye, 7. Vitreomacular traction or epiretinal membrane in the eye, e.g., as evident on biomicroscopy or OCT, which is thought to affect central vision; 8. Preretinal fibrosis involving the macula of the eye, 9. Any history of stage 2 or greater macular hole in the eye, 10. Current iris neovascularization, vitreous hemorrhage, or tractional retinal detachment visible on ophthalmic screening evaluation, 11. History of corneal transplant or corneal dystrophy 12. Any concurrent ocular condition, including: The treating physician's observations may increase the risk to the subject beyond that expected from standard IVT injection procedures; or ocular conditions that may otherwise interfere with the injection procedure of the VEGF antagonist, 13. History of other diseases, metabolic dysfunction, physical examination findings, or clinical laboratory findings that give reasonable suspicion of a disease or condition in which the use of a VEGF antagonist is contraindicated; 14. Previous systemic (IV) administration of any anti-VEGF antibody, 15. Uncontrolled diabetes mellitus defined by hemoglobin A1c (HbA1c) >12%; 16. Uncontrolled blood pressure (defined as systolic >160 mmHg or diastolic >95 mmHg), 17. History of cerebrovascular disease or myocardial infarction within 24 weeks (168 days) of treatment 18. History of renal failure, dialysis, or kidney transplant 19. Known sensitivity to any of the compounds administered in the treatment, and / or 20. Pregnant or lactating women, having or lacking one or more of the following:

[0085] Accordingly, the present invention relates to a subject in need of treatment or prevention of DR and / or DME, 1. Subjects aged ≥ 50 years, 2. Subjects with active subfoveal CNV, 3. Subjects with a best-corrected visual acuity (BCVA) letter score of approximately 78 to 24 on the Early Treatment Diabetic Retinopathy Study (ETDRS). 4. Those with central retinal thickness of ≥ 300 micrometers or ≥ 320 micrometers. 5. Approximately 6 or 7 mm 2 Those with a lesion area of 6. Those with a DRSS score of level 43 or above and level 47 or below, and / or 7. Do you have type 1 or type 2 diabetes? and / or 1. Subjects lacking evidence of macular edema of any cause other than diabetes mellitus in the eye. 2. Those whose IOP is not ≥ 25mmHg in the eye 3. Those who have no history of glaucoma filtration surgery or who are unlikely to require filtration surgery in the future. 4. No evidence of infectious blepharitis, keratitis, scleritis, or conjunctivitis in either eye within 4 weeks (28 days) of treatment. 5. No intraocular inflammation and / or eye infection within 12 weeks (84 days) of treatment. 6. No history of idiopathic or autoimmune uveitis in the eye. 7. Those who do not have vitreomacular traction or epiretinal membrane in their eyes. 8. No preretinal fibrosis involving the retinal macula in the eye. 9. No history of stage 2 or higher macular hole in the eye. 10. No current iris neovascularization, vitreous hemorrhage, and / or tractional retinal detachment. 11. No history of corneal transplantation or corneal dystrophy. 12. Any concurrent ocular condition, including: The treating physician's observations may increase the risk to the subject beyond that expected from standard IVT injection procedures; or who do not have ocular conditions that could otherwise interfere with the injection procedure of a VEGF antagonist; 13. No history of other diseases, metabolic dysfunction, physical examination findings, or clinical laboratory findings that give reasonable suspicion of a disease or condition that contraindicates the use of a VEGF antagonist. 14. Subjects who have not received any systemic (IV) administration of anti-VEGF antibodies in the past. 15. No uncontrolled diabetes mellitus as defined by hemoglobin A1c (HbA1c) >12%. 16. No uncontrolled blood pressure (defined as systolic >160mmHg or diastolic >95mmHg). 17. No history of cerebrovascular disease or myocardial infarction within 24 weeks (168 days) of treatment. 18. No history of renal failure, dialysis, or kidney transplant. 19. Individuals with no known sensitivity to any of the compounds administered in the treatment, and / or 20. A method for treating or preventing DR and / or DME in a subject who is not pregnant or a lactating woman, the method comprising: The method comprises administering to the subject's eye a single initial dose of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, followed by one or more secondary doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more tertiary doses of about 8 mg or more of the VEGF receptor fusion protein, each secondary dose administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose administered about 24 weeks after the immediately preceding dose.

[0086] VEGF antagonists The present invention includes methods of using VEGF antagonists to treat or prevent intraocular neovascular diseases. VEGF antagonists include molecules that interfere with the interaction between VEGF and a native VEGF receptor, such as molecules that bind to VEGF or a VEGF receptor and prevent or otherwise interfere with the interaction between VEGF and a VEGF receptor. Specific exemplary VEGF antagonists include anti-VEGF antibodies, anti-VEGF receptor antibodies, and VEGF receptor fusion proteins. While VEGF receptor fusion proteins, such as aflibercept, are preferred for use in connection with the methods described herein, the scope of the present invention includes such methods in which any of the VEGF antagonists described herein (e.g., scFvs, DARPins, anti-VEGF antibodies) are used in place of such fusion proteins.

[0087] For purposes of this specification, a "VEGF receptor fusion protein" refers to a molecule comprising one or more VEGF receptors or domains thereof fused to another polypeptide, which interferes with the interaction between VEGF and a native VEGF receptor, e.g., when two such fusion polypeptides associate, thereby forming a homodimer or other multimer. Such VEGF receptor fusion proteins are sometimes referred to as "VEGF-Trap" or "VEGF Trap." VEGF receptor fusion proteins within the context of the present disclosure encompassed by this definition include chimeric polypeptides comprising two or more immunoglobulin (Ig)-like domains of a VEGF receptor, such as VEGFR1 (also known as Flt1) and / or VEGFR2 (also known as Flk1 or KDR), and which may also contain a multimerization domain (e.g., an Fc domain).

[0088] An exemplary VEGF receptor fusion protein is a molecule designated VEGF1R2-FcΔC1(a), which is encoded by the nucleic acid sequence of SEQ ID NO: 1, or nucleotides 79 to 1374 or 79 to 1371 thereof.

[0089] VEGF1R2-FcΔC1(a) contains the following three components: (1) a VEGFR1 component comprising amino acids 27 to 129 of SEQ ID NO: 2; (2) a VEGFR2 component comprising amino acids 130 to 231 of SEQ ID NO: 2; (3) A multimer-forming component ("FcΔC1(a)") comprising amino acids 232 to 457 of SEQ ID NO: 2 (the C-terminal amino acid of SEQ ID NO: 2, i.e., K458, may or may not be included in the VEGF receptor fusion protein. See U.S. Patent Nos. 7,396,664 or 7,354,579, which are incorporated herein for all purposes). Note that amino acids 1 to 26 of SEQ ID NO: 2 are a signal sequence.

[0090] When the multimerizing component (MC) of a VEGF receptor fusion protein is derived from an IgG (e.g., IgG1) Fc domain, the MC has as many amino acids as those in amino acids 232 to 457 of SEQ ID NO: 2. Thus, the IgG MC cannot be cleaved to less than 226 amino acids.

[0091] In one embodiment of the invention, the VEGF receptor fusion protein comprises amino acids 27 to 458 or 27 to 457 of SEQ ID NO:2 (eg, in the form of a homodimer). (SEQ ID NO: 1) MVSYWDTGVLLCALLSCLLLTGSSSGSDTGRPFVEMYSEIPEIIIHMTEGRELVIPCRVTS PNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLT HRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRD LKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 2)

[0092] In one embodiment of the invention, the VEGF receptor fusion protein comprises: (1) a second immunoglobin-like (Ig) domain of a first VEGF receptor (e.g., VEGFR1); and (2) the third Ig domain of a second VEGF receptor (e.g., VEGFR2); and (3) optionally, further comprising a fourth Ig domain of a second VEGF receptor (e.g., VEGFR2); and (4) a multimerizing component (e.g., an IgG Fc domain including a hinge domain, a CH2 domain, and a CH3 domain).

[0093] For example, in one embodiment of the present invention, the VEGF receptor fusion protein has the following arrangement of the domains: [VEGFR1 second Ig domain]-[VEGFR2 third Ig domain]-[MC] (e.g., a homodimer thereof), or [2nd Ig domain of VEGFR1]-[3rd Ig domain of VEGFR2]-[4th Ig domain of VEGFR2]-[MC] (e.g., a homodimer thereof).

[0094] The present disclosure also includes within its scope high concentration formulations that include, in place of a VEGF receptor fusion protein, a VEGF binding molecule or an anti-VEGF antibody or antigen-binding fragment thereof, or a biopolymer conjugate thereof (e.g., KSI-301), e.g., Bevacizumab (e.g., at a concentration of about 80-90 or 88 mg / ml), ranibizumab (e.g., at a concentration of about 20-40 mg / ml, e.g., 21-35, 21, or 35 mg / ml); anti-VEGF aptamers, such as pegaptanib (e.g., pegaptanib sodium); Single-chain (e.g., V) antibodies such as brolucizumab (e.g., at a concentration of about 200-400 or 200, 210, 400, or 420 mg / ml) L -V H ) anti-VEGF antibody, an anti-VEGF DARPin, such as abyssiparpegol DARPin (e.g., at a concentration of about 70-140 mg / ml), or bispecific anti-VEGF antibodies that also bind to ANG2, such as RG7716 (faricimab) (e.g., at a concentration of about 100-400, 100, 105, 400, or 420 mg / ml).

[0095] To minimize repetition of embodiments discussed herein, the scope of the present invention is intended to include embodiments in which any of the formulations discussed herein comprises, in place of the VEGF receptor fusion protein, an anti-VEGF antibody or antibody fragment, or other VEGF binding molecule (e.g., substituted with an anti-VEGF DARPin), discussed herein, at any of the concentrations discussed herein. For example, the present invention includes formulations having 35 or 80 mg / ml ranibizumab, a buffer, a heat stabilizer, a viscosity-lowering agent, and a surfactant.

[0096] DARPins are designed ankyrin repeat proteins. They typically contain three to four tightly packed repeats of approximately 33 amino acid residues, each containing a β-turn and two antiparallel α-helices. This rigid framework provides protein stability for target recognition while allowing the display of variable regions, typically containing six amino acid residues per repeat.

[0097] An "anti-VEGF" antibody or antigen-binding fragment of an antibody refers to an antibody or fragment that specifically binds to VEGF.

[0098] Exemplary VEGF receptor fusion proteins include aflibercept (EYLEA®, Regeneron Pharmaceuticals, Inc.) or conbercept (marketed by Chengdu Kanghong Biotechnology Co., Ltd.). See International Patent Application Publication No. WO 2005 / 121176 or WO 2007 / 112675. The terms "aflibercept" and "conbercept" include biosimilar versions thereof. A biosimilar version of a reference product (e.g., aflibercept) generally refers to a product that contains the same amino acid sequence, but includes products that are biosimilar under the U.S. Biologics Price Competition and Innovation Act.

[0099] The present invention also includes embodiments that include administering one or more additional therapeutic agents in addition to a VEGF antagonist, such as administering a second VEGF antagonist (in one or more administrations), an antibiotic, an anesthetic (e.g., a topical anesthetic) to the eye receiving the injection, a nonsteroidal anti-inflammatory drug (NSAID), a steroid (e.g., a corticosteroid, dexamethasone), triamcinolone acetonide (TA), methotrexate, rapamycin, an anti-tumor necrosis factor alpha drug (e.g., infliximab), daxizumab, and / or a complement component (e.g., C3 or C5) inhibitor.

[0100] Pharmaceutical preparations The present invention includes a method in which a VEGF antagonist administered to a subject's eye is contained within a pharmaceutical formulation. The pharmaceutical formulation includes a VEGF antagonist together with a pharmaceutically acceptable carrier. Other agents may be incorporated into the pharmaceutical formulation to provide improvements in transportation, delivery, tolerability, etc. The term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia for use in animals, more particularly for use in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the VEGF antagonist is administered. Many suitable formulations can be found in the formulary known to all pharmacists, Remington's Pharmaceutical Sciences (15th ed., Mack Publishing Company, Easton, PA, 1975), particularly Chapter 87 by Blaug and Seymour.

[0101] Pharmaceutical formulations for use in the methods of the invention can be "highly concentrated." Highly concentrated pharmaceutical formulations of the invention include a VEGF antagonist, e.g., a VEGF receptor fusion protein, at a concentration of at least 41 mg / ml, at least 80 mg / ml, at least 100 mg / ml, at least 125 mg / ml, at least 140 mg / ml, at least 150 mg / ml, at least 175 mg / ml, at least 200 mg / ml, at least 225 mg / ml, at least 250 mg / ml, or at least 275 mg / ml. "Highly concentrated" can refer to a formulation containing a concentration of the VEGF antagonist of about 140 mg / ml to about 160 mg / ml, at least about 140 mg / ml but less than 160 mg / ml, about 41 mg / ml to about 275 mg / ml, about 70 mg / ml to about 75 mg / ml, or about 80 mg / ml to about 250 mg / ml. In some embodiments, the concentration of the VEGF antagonist in the formulation is approximately: 41 mg / ml; 42 mg / ml; 43 mg / ml; 44 mg / ml; 45 mg / ml; 46 mg / ml; 47 mg / ml; 48 mg / ml; 49 mg / ml; 50 mg / ml; 51 mg / ml; 52 mg / ml; 53 mg / ml; 54 mg / ml; 55 mg / ml; 56 mg / ml; 57 mg / ml; 58 mg / ml; 59 mg / ml; 60 mg / ml; 61 mg / ml; 62 mg / ml; 63 mg / ml; 64 mg / ml; 65 mg / ml; 66 mg / ml; 67 mg / ml; 68 mg / ml; 69 mg / ml; 70 mg / ml; 71 mg / ml; 72 mg / ml; 73 mg / ml; 74 mg / ml; 75 mg / ml; mg / ml;76mg / ml;77mg / ml;78mg / ml;79mg / ml;80mg / ml;81mg / ml;82mg / ml;83mg / ml;84mg / ml;85mg / ml;86mg / ml;87mg / ml;88mg / ml;89mg / ml;90mg / ml;91mg / ml;92mg / ml;93mg / ml;94m g / ml;95mg / ml;96mg / ml;97mg / ml;98mg / ml;99mg / ml;100mg / ml;101mg / ml;102mg / ml;103m g / ml;104mg / ml;105mg / ml;106mg / ml;107mg / ml;108mg / ml;109mg / ml;110mg / ml;111mg / ml;112mg / ml;113mg / ml;113.3mg / ml;114mg / ml;114.1mg / ml;114.2mg / ml;114.3mg / ml;114.4mg / ml;114.5mg / ml;114.6mg / ml,114.7mg / ml,114.8mg / ml;114.9mg / ml;115mg / ml;116mg / ml;117mg / ml;118mg / ml;119mg / ml;120mg / ml;121mg / ml;122mg / ml;123mg / ml;124mg / ml;125mg / ml;126mg / ml;127mg / ml;128mg / ml;129mg / ml;130mg / ml;131mg / ml;132mg / ml;133mg / ml;133.3mg / ml;133.4mg / ml,134mg / ml;135mg / ml;136mg / ml;137mg / ml;138mg / ml;139mg / ml;140mg / ml;141mg / ml;142mg / ml;143mg / ml;144mg / ml;145mg / ml;146mg / ml;147mg / ml;148mg / ml;149mg / ml;150mg / ml;151mg / ml;152mg / ml;153mg / ml;154mg / ml;155mg / ml;156mg / ml;157mg / ml;158mg / ml;159mg / ml;160mg / ml;161mg / ml;162mg / ml;163mg / ml;164mg / ml;165mg / ml;166mg / ml;167mg / ml;168mg / ml;169mg / ml;170mg / ml;171mg / ml;172mg / ml;173mg / ml;174mg / ml;175mg / ml;176mg / ml;177mg / ml;178mg / ml;179mg / ml;180mg / ml;181mg / ml;182mg / ml;183mg / ml;184mg / ml;185mg / ml;186mg / ml;187mg / ml;188mg / ml;189mg / ml;190mg / ml;191mg / ml;192mg / ml;193mg / ml;194mg / ml;195mg / ml;196mg / ml;197mg / ml;198mg / ml;199mg / ml;200mg / ml;201mg / ml;202mg / ml;203mg / ml;204mg / ml;205mg / ml;206mg / ml;207mg / ml;208mg / ml;209mg / ml;210mg / ml;211mg / ml;212mg / ml;213mg / ml;214mg / ml;215mg / ml;216mg / ml;217mg / ml;218mg / ml;219mg / ml;220mg / ml;221mg / ml;222mg / ml;223mg / ml;224mg / ml;225 mg / ml;226mg / ml;227mg / ml;228mg / ml;229mg / ml;230mg / ml;231mg / ml;232mg / ml;233mg / ml; 234mg / ml;235mg / ml;236mg / ml;237mg / ml;238mg / ml;239mg / ml;240mg / ml;241mg / ml;242mg / ml;243mg / ml;244mg / ml;245mg / ml;246mg / ml;247mg / ml;248mg / ml;249mg / ml;250mg / ml;251 mg / ml;252mg / ml;253mg / ml;254mg / ml;255mg / ml;256mg / ml;257mg / ml;258mg / ml;259mg / ml; 260mg / ml;261mg / ml;262mg / ml;263mg / ml;264mg / ml;265mg / ml;266mg / ml;267mg / ml;268mg / 269 ​​mg / ml; 270 mg / ml; 271 mg / ml; 272 mg / ml; 273 mg / ml; 274 mg / ml; or 275 mg / ml. Other concentrations of the VEGF antagonist are contemplated herein, so long as the concentrations function in accordance with the embodiments herein.

[0102] In one embodiment of the invention, the pharmaceutical formulation for use in the methods of the invention is concentrated to contain about 4, 6, 8, 10, 12, 14, 16, 18, or 20 mg of VEGF receptor fusion protein (e.g., aflibercept), or at any of the acceptable doses thereof discussed herein, in a volume of about 100 μl or less, about 75 μl or less, or about 70 μl or less, such as about 50 μl, 51 μl, 52 μl, 53 μl, 54 μl, 55 μl, 56 μl, 57 μl, 58 μl, 59 μl, 60 μl, 61 μl, 62 μl, 63 μl, 64 μl, 65 μl, 66 μl, 67 μl, 68 μl, 69 μl, 70 μl, 71 μl, 72 μl, 73 μl, 74 μl, 75 μl, 76 μl, 77 μl, 78 μl, 79 μl, 80 μl, 81 μl, 82 μl, 83 μl, 84 μl, 85 μl, 86 μl, 87 μl, 88 μl, 89 μl, 90 μl, 91 μl, 92 μl, 93 μl, 94 μl, 95 μl, 96 μl, 97 μl, 98 μl, 99 μl, or 100 μl of the protein.

[0103] The present invention includes methods of using any of the formulations described in the "Exemplary Formulations" section herein (as discussed herein), but substituting the concentrations of VEGF receptor fusion proteins (e.g., aflibercept) with those described in this section ("VEGF Receptor Fusion Proteins and Other VEGF Inhibitors").

[0104] A buffer for use in the pharmaceutical formulations herein that can be used in the methods of the present invention refers to a solution that resists pH changes due to the use of an acid-base complex. The buffer can maintain a pH within the range of about 5.0 to about 6.8, more typically about 5.8 to about 6.5, and most typically about 6.0 to about 6.5. In some cases, the pH of the formulations of the present invention is about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, or about 6.8. Examples of buffers for inclusion in the formulations herein include histidine-based buffers, such as histidine and histidine hydrochloride or histidine acetate. Alternatively, the buffer included in the formulation herein may be a phosphate-based buffer, such as sodium phosphate; an acetate-based buffer, such as sodium acetate or acetic acid; or a citrate-based buffer, such as sodium citrate or citric acid. It is also recognized that the buffer may be a mixture of the above, so long as the buffer functions to buffer the formulation within the pH ranges described above. In some cases, the buffer is about 5 mM to about 25 mM, or more typically about 5 mM to about 15 mM. The buffer may be about 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM.

[0105] In one embodiment of the present invention, the histidine-based buffer is prepared using histidine and histidine monohydrochloride.

[0106] As used herein, surfactant refers to a component that protects higher concentrations of VEGF antagonists, such as VEGF receptor fusion proteins, from various surface- and interface-induced stresses. Thus, surfactants can be used to limit or minimize aggregation of VEGF receptor fusion proteins and promote protein solubility. Suitable surfactants herein are known to be nonionic and may include surfactants with polyoxyethylene moieties. Exemplary surfactants in this category include polysorbate 20, polysorbate 80, poloxamer 188, polyethylene glycol 3350, and mixtures thereof. The surfactant in the formulation may be present at about 0.02% to about 0.1% weight per volume (w / v), more typically about 0.02% to about 0.04% (w / v). In some cases, the surfactant is about 0.02% (w / v), about 0.03% (w / v), about 0.04% (w / v), about 0.05% (w / v), about 0.06% (w / v), about 0.07% (w / v), about 0.08% (w / v), about 0.09% (w / v), or about 0.1% (w / v).

[0107] The thermostabilizer for use in the pharmaceutical preparations described herein refers to a component that provides thermal stability to the VEGF antagonist, such as a VEGF receptor fusion protein, against thermal denaturation and protects the VEGF receptor fusion protein from losing its potency or activity. Suitable thermostabilizers include sugars, such as sucrose, trehalose, sorbitol, or mannitol, or amino acids such as L-proline, L-arginine (e.g., L-arginine monohydrochloride), or taurine. Additionally, thermostabilizers may include substituted acrylamides or propanesulfonic acids, or may be compounds such as glycerol.

[0108] In some instances, pharmaceutical formulations for use in the methods herein include a sugar and taurine, a sugar and an amino acid, a sugar and propanesulfonic acid, a sugar and taurine, glycerol and taurine, glycerol and propanesulfonic acid, an amino acid and taurine, or both an amino acid and propanesulfonic acid. Further, the formulation may include a sugar, taurine, and propanesulfonic acid, glycerol, taurine, and propanesulfonic acid, and L-proline, taurine, and propanesulfonic acid.

[0109] Embodiments herein may have thermal stabilizers present singly, each independently at a concentration of about 2% (w / v) to about 10% (w / v), or 4% (w / v) to about 10% (w / v), or about 4% (w / v) to about 9% (w / v), or about 5% (w / v) to about 8% (w / v), or at a combined total concentration. The thermal stabilizers in the formulation may be at a concentration of about 2% (w / v), about 2.5% (w / v), about 3% (w / v), about 4% (w / v), about 5% (w / v), about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), about 10% (w / v), or about 20% (w / v).

[0110] With respect to taurine and propanesulfonic acid, in one embodiment of the present invention, these heat stabilizers may be present in the formulation at about 25 mM to about 100 mM, more typically about 50 mM to about 75 mM (relative to other heat stabilizers).

[0111] Viscosity-lowering agents are typically used to reduce or prevent protein aggregation. Viscosity-lowering agents included herein include sodium chloride, magnesium chloride, D-arginine or L-arginine (e.g., L-arginine monohydrochloride), lysine, or mixtures thereof. When present herein, the viscosity-lowering agent may be present at about 10 mM to about 100 mM, more typically at about 30 mM to about 75 mM, and even more typically at about 40 mM to about 70 mM. In some cases, the viscosity-lowering agent is present at about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM.

[0112] Pharmaceutical formulations for use in the methods described herein can also have a pharmaceutically acceptable viscosity for ocular administration, e.g., intravitreal injection. Viscosity generally refers to a measure of a fluid's resistance to being deformed by either shear or tensile stress (typically measured, e.g., by techniques known in the art, a viscometer, or a rheometer). Typical viscosities of formulations for use in the methods described herein are about 5.0 cP (centipoise) to about 15 cP, about 11 cP to about 14 cP, about 12 cP to about 15 cP, or about 11 cP to about 12 cP. Thus, the viscosity of the formulations herein can be about 5.0 cP, about 6.0, about 7.1 cP, about 7.2 cP, about 7.3 cP, about 7.4 cP, about 7.5 cP, about 7.6 cP, about 10 cP, about 10.5 cP, about 11.0 cP, about 11.5 cP, about 12.0 cP, about 12.5 cP, about 13.0 cP, about 13.5 cP, about 14.0 cP, about 14.5 cP, or about 15.0 cP (e.g., when measured at 20°C).

[0113] Various embodiments herein do not require the inclusion of inorganic salts or other viscosity-lowering agents to maintain these highly useful viscosities. Typically, highly concentrated protein solutions require viscosity-lowering agents to avoid protein aggregation and high viscosity, which can complicate intravitreal injection of the formulation and reduce the efficacy of the VEGF receptor fusion protein. Accordingly, embodiments herein include methods using formulations that are substantially free of, or do not contain, sodium chloride (NaCl), magnesium chloride (MgCl), D-arginine or L-arginine (e.g., L-arginine hydrochloride), lysine, or other viscosity-lowering agents.

[0114] Osmolality is an important characteristic of injectable pharmaceutical formulations for use in the methods of the present invention. It is desirable for the formulation to match physiological osmolality conditions. Furthermore, osmolality determines the soluble content of the solution. In one embodiment of the present invention, the osmolality of a formulation for use in the methods of the present invention is about 506 mmol / Kg or less, or about 250 to about 506 mmol / Kg, e.g., about 250, 260, 270, 280, 290, 299, 300, 310, 314, 315, 316, 324, 343, 346, 349, 369, 384, 403, 426, 430, or 506 mmol / Kg. In one embodiment of the present invention, the osmolality is less than about 250 mmol / Kg.

[0115] Exemplary pharmaceutical formulations for use in the methods of the present invention include: Formulation A: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation B: 80 mg / ml aflibercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation C: 80 mg / ml aflibercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation D: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2; Formulation E: 80 mg / ml aflibercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation F: 80 mg / ml aflibercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation G: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation H: 80 mg / ml aflibercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation I: 80 mg / ml aflibercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation J: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation K: 80 mg / ml aflibercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation L: 80 mg / ml aflibercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation M: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation N: 150 mg / ml aflibercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation O: 150 mg / ml aflibercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation P: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2; Formulation Q: 150 mg / ml aflibercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation R: 150 mg / ml aflibercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation S: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally, specifically excluding a viscosity-lowering agent; Formulation T: 150 mg / ml aflibercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2 (e.g., 6.2), optionally excluding a viscosity-lowering agent; Formulation U: 150 mg / ml aflibercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation V: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally, specifically excluding a viscosity-lowering agent; Formulation W: 150 mg / ml aflibercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation X: 150 mg / ml aflibercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation Y: 80 mg / ml conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation Z: 80 mg / ml conbercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation AA: 80 mg / ml conbercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation BB: 80 mg / ml conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2; Formulation CC: 80 mg / ml conbercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation DD: 80 mg / ml conbercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation EE: 80 mg / ml conbercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally, specifically excluding a viscosity-lowering agent; Formulation FF: 80 mg / ml conbercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding viscosity-reducing agents; Formulation GG: 80 mg / ml conbercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation HH: 80 mg / ml conbercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation II: 80 mg / ml conbercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding a viscosity-lowering agent; Formulation JJ: 80 mg / ml conbercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation KK: 150 mg / ml conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation LL: 150 mg / ml conbercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation MM: 150 mg / ml conbercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8-6.2; Formulation NN: 150 mg / ml conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2; Formulation 00: 150 mg / ml conbercept, 10 mM phosphate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation PP: 150 mg / ml conbercept, 10 mM citrate buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8-6.2; Formulation QQ: 150 mg / ml conbercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding viscosity-reducing agents; Formulation RR: 150 mg / ml conbercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding viscosity-reducing agents; Formulation SS: 150 mg / ml conbercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation TT: 150 mg / ml conbercept, 10 mM histidine-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation UU: 150 mg / ml conbercept, 10 mM phosphate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation VV: 150 mg / ml conbercept, 10 mM citrate buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8-6.2, optionally excluding viscosity-lowering agents; Formulation WW: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM taurine, pH 5.8; Formulation XX: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 4% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM arginine (e.g., arginine hydrochloride), pH 5.8; Formulation YY: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM taurine, pH 5.8; Formulation ZZ: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM arginine (e.g., arginine hydrochloride), pH 5.8; Formulation AAA: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8; Formulation BBB: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8, Formulation CCC: 80, 100, 120, or 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM arginine (e.g., arginine hydrochloride), pH 5.8; Formulation DDD: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM histidine-based buffer, 4% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8; Formulation EEE: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, optionally without a heat stabilizer, pH 5.8; Formulation FFF: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM sodium phosphate, 5% (w / v) sucrose, and 0.03% polysorbate 20, pH 6.2, Formulation GGG: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium sulfate, Formulation HHH: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium sulfate, Formulation III: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 40 mM sodium citrate, Formulation JJJ: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM glycine, Formulation KKK: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium chloride, Formulation LLL: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM lysine, Formulation MMM: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium aspartate, Formulation NNN: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium glutamate, Formulation 0000: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium citrate, 50 mM arginine (e.g., arginine hydrochloride), Formulation PPP: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM glycine, 50 mM arginine (e.g., arginine hydrochloride), Formulation QQQ: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium citrate, 50 mM arginine (e.g., arginine hydrochloride), Formulation RRR: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 50 mM sodium citrate, 50 mM arginine (e.g., arginine hydrochloride), Formulation SSS: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 10 mM L-arginine (e.g., L-arginine hydrochloride), Formulation TTT: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine, pH 5.8, 5% sucrose, 0.03% polysorbate 20, 100 mM L-arginine (e.g., L-arginine hydrochloride), Formulation UUU: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation VVV: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation WWW: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation XXX: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation YYY: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation ZZZ: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2, Formulation AAAA: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, 50 mM NaCl, pH 6.2, Formulation BBBB: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, 50 mM NaCl, pH 6.2, Formulation CCCC: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM sodium phosphate, 5% sucrose, 40 mM sodium chloride, 0.03% PS20, pH 6.2, Formulation DDDD: 80 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine (e.g., L-arginine hydrochloride), pH 5.8, Formulation EEEE: 120.0 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±12 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine hydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1), Formulation FFFF: 113.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., 102-125 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1), Formulation GGGG: 114.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., 103-126 mg / ml), 10 mM histidine-based buffer, such as one containing histidine and histidine-HCl (e.g., ±1 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1), Formulation HHHH: 100.0 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±10 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1), Formulation III: 133.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±13 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1), Formulation JJJJ: 150 mg / ml aflibercept (e.g., aflibercept) (e.g., ±15 mg / ml), 10 mM sodium phosphate, 8% (w / v) sucrose (e.g., ±0.8%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., arginine hydrochloride), pH 6.2 (e.g., 6.0-6.4 or 5.9-6.5), Formulation KKKK: 114.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±14 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0116] See International Patent Application Publication No. WO 2019 / 217927.

[0117] In one embodiment of the present invention, at least 8 mg of a VEGF receptor fusion protein, preferably aflibercept, is administered in an aqueous pharmaceutical formulation containing two polypeptides, each comprising the second immunoglobulin-like (Ig) domain of VEGFR1, the third Ig domain of VEGFR2, and a multimerization component (e.g., amino acids 27-457 of SEQ ID NO: 2), at a concentration of at least about 100 mg / ml; about 5% sucrose; L-arginine (e.g., L-arginine monohydrochloride); a histidine-based buffer (e.g., containing histidine hydrochloride); and about 0.03% surfactant, with a pH of about 5.0 to about 6.8 (e.g., 5.8 to 6.5, e.g., 5.8). The formulation is preferably suitable for intravitreal administration. Other components that may be included include sodium sulfate, sodium thiocyanate, glycine, NaCl, sodium aspartate, and / or sodium glutamate. In one embodiment of the present invention, the VEGF receptor fusion protein is at a concentration of about 100 mg / ml, about 111.5 mg / ml, about 112.0 mg / ml, about 113.3 mg / ml, about 114.3 mg / ml, about 115.6 mg / ml, about 116.3 mg / ml, about 120 mg / ml, about 133 mg / ml, about 140 mg / ml, about 150 mg / ml, about 200 mg / ml, or about 250 mg / ml. The formulation may be characterized by (i) an osmolality of about 299 to about 506 mmol / kg and / or (ii) a viscosity of about 6 to 15 cP at 20°C. The surfactant may be a non-ionic surfactant such as polysorbate 20, polysorbate 80, poloxamer 188, polyethylene glycol 3350, or a mixture thereof. The histidine-based buffer may be at a concentration of about 10 mM to 20 mM. In one embodiment of the present invention, the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification, and / or about 6% or less high molecular weight species after storage at about 2-8°C for about 24 months.

[0118] In one embodiment of the present invention, at least 8 mg of VEGF receptor fusion protein is present in the aqueous pharmaceutical formulation when administered, comprising at least about 100 mg / ml of VEGF receptor fusion protein comprising two polypeptides, each comprising a second immunoglobulin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, and a multimerization component (e.g., aflibercept); about 10-100 mM L-arginine, sucrose, a histidine-based buffer, and a surfactant, wherein the formulation has a pH of about 5.0 to about 6.8, and the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification and / or about 6% or less high molecular weight species after storage for about 24 months at about 2-8°C.

[0119] In one embodiment of the present invention, the pharmaceutical formulation comprises: ≥ about 100 mg / ml of a VEGF receptor fusion protein (e.g., aflibercept), a histidine-based buffer, and L-arginine; Approximately 140 mg / ml aflibercept, 20 mM histidine-based buffer, 5% sucrose, 0.03% polysorbate 20, 10 mM L-arginine, pH 5.8, Approximately 150±15 mg / ml aflibercept, 10 mM phosphate buffer, 8±0.8% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.9-6.5. Approximately 103-126 mg / ml aflibercept, 10 ± 1 mM histidine-based buffer, 5 ± 0.5% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20, and 50 ± 5 mM L-arginine, pH 5.5-6.1; Approximately 140 mg / ml aflibercept, 10 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8; Approximately 114.3 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8; ≥ about 100 mg / ml aflibercept, a histidine-based buffer, and L-arginine; aflibercept of ≥ about 100 mg / ml at a pH of about 5.8, wherein the formulation forms <3% HMW aggregates after 2 months of incubation at 5°C; Approximately 114.3 mg / mL aflibercept, 10 mM to 50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, pH 5.8, or Approximately 114.3 mg / mL aflibercept, 10 mM His / His-HCl buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, pH 5.8.

[0120] In one embodiment of the present invention, ≥ 8 mg of VEGF receptor fusion protein is in the aqueous pharmaceutical formulation when administered; aflibercept at a concentration of at least about 100 mg / ml (e.g., about 111.5 mg / ml, 112.0 mg / ml, 113.3 mg / ml, about 114.3 mg / ml, about 115.6 mg / ml, or about 116.3 mg / ml); a heat stabilizer that is a sugar, an amino acid, sucrose, mannitol, sorbitol, trehalose, L-proline, glycine, glycerol, taurine, or propanesulfonic acid (e.g., about 2% (w / v) to about 10% (w / v), e.g., 5% (w / v)); a buffer solution which is a histidine-based buffer solution, a phosphate-based buffer solution, an acetate-based buffer solution (e.g., at a concentration of about 5 to 25 mM, e.g., 10 mM or 20 mM), or a citrate-based buffer solution; Non-ionic surfactants, such as polyoxyethylene surfactants, polysorbate 20, polysorbate 80, poloxamer 188, or polyethylene glycol 3350 (e.g., at a concentration of about 0.02% to about 0.1% (w / v), e.g., 0.03% (w / v)), and a viscosity-lowering agent (e.g., at a concentration of about 10-100 mM, e.g., 50 mM) that is NaCl, MgCl, D-arginine, L-arginine, or L-lysine; The formulation has a pH of about 5.0 to about 6.8 (eg, 5.0 to 6.0 or 5.8).

[0121] In one embodiment of the present invention, the concentration of aflibercept in the aqueous pharmaceutical formulation is about 100 mg / ml, 101 mg / ml; 102 mg / ml; 103 mg / ml; 104 mg / ml; 105 mg / ml; 106 mg / ml; 107 mg / ml; 108 mg / ml; 109 mg / ml; 110 mg / ml; 111 mg / ml; 112 mg / ml; 113 mg / ml; 113.3 mg / ml; 114 mg / ml; 114.1 mg / ml; 114.2 mg / ml; 114.3 mg / ml; 114.4 mg / ml; 114.5 mg / ml; 114.6 mg / ml, 114.7 mg / ml, 114.8 mg / ml; 114.9 mg / ml; 115 mg / ml; 116 mg / ml; 117 mg / ml; 118 mg / ml; 119 mg / ml; 120 mg / ml; 121 mg / ml; 122 mg / ml; 123 mg / ml; 124 mg / ml; 125 mg / ml; 126 mg / ml; 127 mg / ml; 128 mg / ml; 129 mg / ml; 130 mg / ml; 131 mg / ml; 132 mg / ml; 133 mg / ml; 133.3 mg / ml; 133.4 mg / ml, 134 mg / ml; 135 mg / ml; 136 mg / ml; 137 mg / ml; 138 mg / ml; 139 mg / ml; 140 mg / ml; 141 mg / ml; 142 mg / ml; 143 mg / ml; 144 mg / ml; 145 mg / ml; 146 mg / ml; 147 mg / ml; 148 mg / ml; 149 mg / ml; 150 mg / ml; 151 mg / ml; 152 mg / ml; 153 mg / ml; 154 mg / ml; 155 mg / ml; 156 mg / ml; 157 mg / ml; 158 mg / ml; 159 mg / ml; 160 mg / ml; 161 mg / ml; 162 mg / ml; 163 mg / ml; 164 mg / ml; 165 mg / ml; 166 mg / ml; 167 mg / ml; 168 mg / ml; 169 mg / ml; 170 mg / ml; 171 mg / ml; 172 mg / ml; 173 mg / ml; 174 mg / ml; 175 mg / ml; 176 mg / ml; 177 mg / ml; 178 mg / ml; 179 mg / ml; 180 mg / ml; 181 mg / ml; 182 mg / ml; 183 mg / ml; 184 mg / ml; 185 mg / ml; 186 mg / ml; 187 mg / ml; 188 mg / ml; 189 mg / ml; 190 mg / ml; 191 mg / ml;192 mg / ml; 193 mg / ml; 194 mg / ml; 195 mg / ml; 196 mg / ml; 197 mg / ml; 198 mg / ml; 199 mg / ml; 200 mg / ml; 201 mg / ml; 202 mg / ml; 203 mg / ml; 204 mg / ml; 205 mg / ml; 206 mg / ml; 207 mg / ml; 208 mg / ml; 209 mg / ml; 210 mg / ml; 211 mg / ml; 212 mg / ml; 213 mg / ml; 214 mg / ml; 215 mg / ml; 216 mg / ml; 217 mg / ml; 218 mg / ml; 219 mg / ml; 220 mg / ml; 221 mg / ml; 222 mg / ml; 223 mg / ml; 224 mg / ml; 225 mg / ml; 226 mg / ml; 227 mg / ml; 228 mg / ml; 229 mg / ml; 230 mg / ml; 231 mg / ml; 232 mg / ml; 233 mg / ml; 234 mg / ml; 235 mg / ml; 236 mg / ml; 237 mg / ml; 238 mg / ml; 239 mg / ml; 240 mg / ml; 241 mg / ml; 242 mg / ml; 243 mg / ml; 244 mg / ml; 245 mg / ml; 246 mg / ml; 247 mg / ml; 248 mg / ml; 249 mg / ml; 250 mg / ml; 251 mg / ml; 252 mg / ml; 253 mg / ml; 254 mg / ml; 255 mg / ml; 256 mg / ml; 257 mg / ml; 258 mg / ml; 259 mg / ml; 260 mg / ml; 261 mg / ml; 262 mg / ml; 263 mg / ml; 264 mg / ml; 265 mg / ml; 266 mg / ml; 267 mg / ml; 268 mg / ml; 269 mg / ml; 270 mg / ml; 271 mg / ml; 272 mg / ml; 273 mg / ml; 274 mg / ml, or 275 mg / ml.

[0122] In one embodiment of the present invention, the aqueous pharmaceutical formulation comprises aflibercept at a concentration of at least about 100 mg / ml, sucrose, mannitol, sorbitol, trehalose, a histidine-based buffer, polysorbate 20 or polysorbate 80, and L-arginine, and has a pH of about 5.0 to about 6.8, wherein the aflibercept has less than about 3.5% high molecular weight species immediately after production and purification, and / or about 6% or less high molecular weight species after storage at about 2 to 8°C for about 24 months.

[0123] In one embodiment of the present invention, the sucrose, mannitol, sorbitol, or trehalose is at a concentration of about 2 to 10% (w / v), the L-arginine is at a concentration of about 10 to 100 mM, the polysorbate 20 or polysorbate 80 is at a concentration of about 0.02 to 0.1% (w / v), and the histidine-based buffer is at a concentration of about 5 to 25 mM, and the pH is about 5.0 to about 6.8.

[0124] Treatment and Administration The present invention provides methods for treating intraocular neovascular diseases (e.g., nAMD, DR and / or DME) by administering an initial loading dose (e.g., 2 mg or more, 4 mg or more, or preferably about 8 mg or more of a VEGF antagonist or inhibitor, e.g., a VEGF receptor fusion protein such as aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks), followed by sequential boost doses every 24 weeks. For example, the present invention provides a method for treating or preventing intraocular neovascular diseases, such as neovascular age-related macular degeneration (nAMD), diabetic macular edema (DME), and / or diabetic retinopathy (DR), by sequentially administering one or more doses (e.g., 3, 4, or 5 doses) of about 8 mg or more of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) about every 2-4 weeks or 3-5 weeks, e.g., monthly (or about every 28 days, 28±5 days, or about every 4 weeks), followed by one or more doses of about 8 mg or more of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) every 24 weeks (or about once every 6 months, or about quarterly, or about every 168 days). A dosing regimen comprising a tertiary administration interval of about 24 weeks may be referred to herein as a 24-week dosing regimen, or 8q24, or HDq24.

[0125] Additionally, the present invention includes methods for treating intraocular neovascular diseases (e.g., nAMD, DR and / or DME) by administering ≥ 8 mg of a VEGF receptor fusion protein, preferably aflibercept, one or more times about every 24 weeks, with the first 3, 4, or 5 administrations about every 4 weeks, followed by administrations about every 24 weeks.

[0126] In one embodiment of the present invention, the subject begins receiving a monthly loading dose of ≥ 8 mg followed by a maintenance dose of ≥ 8 mg approximately every 24 weeks with no intervening doses. The subject enters the maintenance dose phase immediately / immediately after the loading dose phase. In one embodiment of the present invention, the subject continues to receive ≥ 8 mg doses for 24 weeks with no intervening doses.

[0127] For example, the present invention also provides a method for treating an intraocular neovascular disease (preferably, nAMD, DME, or DR), comprising: about ≧8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, e.g., about 50 μl, 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 74 μl; 75 μl 96 μl; 97 μl; 98 μl; 99 μl, or 100 μl) once about every 24 weeks.

[0128] In one embodiment of the present invention, the subject does not undergo a dosing regimen change (DRM) or complete treatment for at least 1, 2, 3, 4, or 5 years.

[0129] The present invention also provides methods for improving vision in subjects with type 1 diabetes or type 2 diabetes (e.g., subjects with neovascular age-related macular degeneration (nAMD), diabetic macular edema, or diabetic retinopathy) by sequentially administering one or more doses (e.g., 3, 4, or 5 doses) about monthly (or about every 28 days, 28±5 days, or about every 4 weeks), followed by one or more doses every 24 weeks.

[0130] The terms "initial administration", "secondary administration", and "tertiary administration" refer to the time sequence of administration of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept). Thus, an "initial administration" refers to the dose administered at the beginning of a treatment regimen (also referred to as a "baseline administration"); a "secondary administration" refers to the dose administered after the initial administration; and a "tertiary administration" refers to the dose administered after the secondary administration. The initial administration, the secondary administration, and the tertiary administration can all contain the same amount of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), but may generally differ from each other in terms of the number of administrations. However, in certain embodiments, the amount of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) contained in the initial administration, the secondary administration, and / or the tertiary administration will vary from each other during the course of treatment (e.g., adjusted upward or downward as appropriate).

[0131] Thus, the dosing regimen of the present invention can be expressed as follows: A method for treating an intraocular neovascular disease (e.g., nAMD, DME, or DR) in a subject in need thereof, comprising administering to the subject (e.g., intravitreally) A single initial dose of about ≥ 8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, e.g., about 50 μl, 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 74 μl; 75 μl; 76 μl; 77 μl; 78 μl; 79 μl; 80 μl; 81 μl; 82 μl; 83 μl; 84 μl; 85 μl; 86 μl; 87 μl; 88 μl; 89 μl; 90 μl; 91 μl; 92 μl; 93 μl; 94 μl; 95 μl; 96 μl; 97 μl; 98 μl; 99 μl, or 100 μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), then one or more (e.g., two, or three, or four, preferably two) secondary administrations of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), then administering one or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept); each secondary administration is administered about 2 to 4 weeks (preferably about 4 weeks) after the immediately preceding administration; and wherein each tertiary administration is administered about 24 weeks after the immediately preceding administration.

[0132] The present invention also relates to a method for treating intraocular neovascular diseases (e.g., nAMD, DR, or DME), comprising administering to a subject in need thereof about ≧8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, e.g., about 50 μl, 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 96μl; 97μl; 98μl; 99μl; or 100μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) on a PRN basis.

[0133] As-needed (PRN) treatment protocols require that the interval between physician visits remain fixed (e.g., once every 2, 3, 4, 8, 12, 16, 20, or 24 weeks) and that the decision to administer VEGF receptor fusion protein injections is based on anatomical findings at each visit. A capped PRN dosing regimen is a PRN method in which the subject must be treated a specified minimum number of times, for example, at least once every 2, 3, 4, or 6 months.

[0134] The Treat & Extend (T&E) regimen involves adjusting the time interval between physician visits based on the patient's clinical progress; for example, if the subject shows no signs of active disease (e.g., the macula remains dry and there is no leakage), the interval between one or more subsequent visits can be extended, whereas if fluid accumulation occurs, the interval between subsequent visits is shortened. In T&E, injections of the VEGF receptor fusion protein are administered at each visit, and only the patient's current clinical status affects the duration of the interval between subsequent injections.

[0135] The present invention includes embodiments in which, at any point during an HDq24 treatment regimen, a patient can switch to a PRN, capped PRN, or T&E regimen. PRN, capped PRN, and / or T&E may be continued indefinitely or may be stopped at any point, after which the HDq24 regimen is resumed at any stage. Any HDq24 regimen can precede or follow a period of PRN, capped PRN, and / or T&E.

[0136] The present invention includes methods of administering to a subject one or more unscheduled additional doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) in addition to any of the scheduled initial, secondary, and / or tertiary doses. Such doses are typically administered at the discretion of the treating physician, depending on the particular needs of the subject.

[0137] Thus, the invention includes methods comprising administering the required dose of the HDq24 regimen, wherein each tertiary administration is administered about 24 weeks after the immediately preceding administration, and wherein the treatment interval between two tertiary administrations is extended (e.g., from about 4, 8, 12, 16, or 20 weeks to about 24 weeks), for example, until signs of disease activity return or vision worsens, and then either continuing administration at the last tertiary interval used or continuing administration at the penultimate tertiary interval used.

[0138] The invention includes methods comprising administering the required dose of an HDq24 regimen, wherein the treatment interval between any two tertiary administrations is shortened (e.g., from about 24 weeks to about 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 weeks), e.g., until signs of disease activity decrease or visual acuity improves (e.g., BCVA stabilizes or improves, and / or CRT stabilizes or decreases), and then optionally, the interval between administrations can be extended, e.g., returned to a longer interval.

[0139] For example, in one embodiment of the present invention, during a dosing phase of, e.g., 12, 16, 20, or 24 weeks, the interval between doses can be, e.g., BCVA loss of <5 letters, e.g., from week 12, and / or If CRT is <300 μm on SD-OCT (or <320 μm on Spectralis SD-OCT), it can be extended in 4-week increments (e.g., from 20 weeks to 24 weeks) if necessary. In one embodiment of the invention, subjects receive an initial dose, a secondary dose, then a tertiary interval of 20 weeks, and then after about one year, the tertiary interval is extended to about 24 weeks.

[0140] In one embodiment of the present invention, the methods for treating intraocular neovascular diseases such as nAMD, DR, or DME described herein include assessing BCVA and / or CRT and extending the interval as described if one or both criteria are met.

[0141] For example, in one embodiment of the present invention, during the administration phase, e.g., at week 24, the interval between administrations may be, e.g., loss of more than 5 or 10 letters in BCVA (ETDRS or equivalent Snellen visual acuity) (e.g., compared to the BCVA observed at approximately 12 or 16 weeks after the start of treatment), e.g., due to or associated with persistence or worsening of nAMD, DR and / or DME; and / or If an increase in CRT of more than 25 or 50 micrometers is observed (e.g., compared to the CRT observed approximately 12 weeks after initiation of treatment), the treatment period can be shortened (e.g., from 24 weeks to 20, 16, 12, or 8 weeks).

[0142] In one embodiment of the present invention, the interval between tertiary administrations is increased or decreased in 4-week increments. The decision to increase or decrease the tertiary administration interval can be made in one or more visits to the treating physician.

[0143] In one embodiment of the present invention, in a subject receiving an HDq24 regimen, if the criteria for shortening the interval between doses are met, the interval between doses is reduced to 20 weeks. In one embodiment of the present invention, the interval is not shortened to a period shorter than 8 weeks. In one embodiment of the present invention, the method of treating intraocular neovascular diseases such as nAMD, DR, or DME described herein includes assessing BCVA and / or CRT and shortening the interval as described if one or both of the criteria are met.

[0144] Please refer to FIG. 3 or FIG.

[0145] Dosing "every 24 weeks" refers to dosing about every 6 months, about every 168 days (±5 days), or about quarterly, or about twice a year.

[0146] Administration "every month" or after "one month" means administration after about 28 days, about 4 weeks, or about 28±5 days, which can include up to 5 weeks±5 days. Administration "every four weeks" or after "four weeks" means administration after about 28 days (±5 days), about one month, or about 28 (±5 days), which can include up to every 5 weeks (±5 days).

[0147] Administration "every 2-4 weeks" or after "2-4 weeks" refers to administration after about 2 weeks (±5 days), 3 weeks (±5 days), or 4 weeks (±5 days). Administration "every 8 weeks" or after "8 weeks" refers to administration after about 2 months (±5 days) or about 56 days (±5 days).

[0148] Dosing "every 12 weeks" or after "12 weeks" means dosing after about 3 months, about 84 days (± 5 days), about 90 days (± 5 days), or about 84 (± 5 days). Dosing "every 16 weeks" or after "16 weeks" refers to dosing after about 4 months or about 112 days (± 5 days).

[0149] Administration "every 12 to 20 weeks" or after "12 to 20 weeks" refers to administration after 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks (±5 days), preferably after about 12 to 16 weeks (±5 days), about 12 weeks (±5 days), about 16 weeks (±5 days), or about 20 weeks (±5 days).

[0150] Administration "every 12 to 20 weeks" refers to administration after about 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks (±5 days), preferably after about 12 to 16 weeks (±5 days), about 12 weeks (±5 days), about 16 weeks (±5 days), or about 20 weeks (±5 days).

[0151] A dose of >= 8 mg includes a dose of about 8 mg, or a dose greater than 8 mg, such as about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg.

[0152] Any dosing frequency specified herein may be expressed in embodiments of the invention as "±5 days" of that particular frequency (e.g., if "24 weeks" is stated, the invention also includes embodiments such as 24 weeks ±5 days). The term ±5 days includes ±1, ±2, ±3, ±4, and / or ±5 days.

[0153] By "sequentially administered" is meant that each dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) is administered to the subject's eye at different times, such as on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). The present invention includes methods that include sequentially administering to the subject's eye a single initial dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by one or more secondary doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by one or more tertiary doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept).

[0154] An effective or therapeutically effective dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) for treating or preventing an intraocular neovascular disease refers to an amount of the VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) sufficient to alleviate one or more signs and / or symptoms of the disease or condition in a treated subject, either by inducing regression or elimination of the signs and / or symptoms or by inhibiting the progression of the signs and / or symptoms. In one embodiment of the present invention, the effective or therapeutically effective dose of the VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) is about ≥ 8 mg administered three times monthly, then once every 24 weeks. In one embodiment of the present invention, a reduction in signs and / or symptoms may be achieved, for example, at one year, by achieving a gain (compared to baseline) of ≥ 5, 10, or 15 letters of BCVA (e.g., an improvement of ≥ 5 letters in nAMD subjects and / or an improvement of 8-14 letters in DME patients / subjects), achieving a BCVA of ≥ 69 letters, achieving absence of fluid at the fovea, a reduction in central retinal thickness (CRT) of about 150 micrometers or more (e.g., less than 300 micrometers in nAMD subjects / patients and / or a reduction by at least about 200 micrometers in DR or RVO patients / subjects), or achieving a normal CRT (e.g., about 300 micrometers or less), and / or no leakage on fluorescein angiography.

[0155] Baseline values ​​refer to values ​​before treatment begins (before administration).

[0156] "Intraocular neovascular disease" means any disease within the eye caused by or associated with the growth or proliferation of blood vessels or with vascular leakage. Non-limiting examples of intraocular neovascular diseases treatable or preventable using the methods of the present invention include: Age-related macular degeneration (neovascular age-related macular degeneration (nAMD)), ·Macular edema (ME), Macular edema after retinal vein occlusion (ME-RVO), Retinal vein occlusion (RVO), Central retinal vein occlusion (CRVO), Branch retinal vein occlusion (BRVO), ·Diabetic macular edema (DME), ·Choroidal neovascularization (CNV), ·iris neovascularization, Neovascular glaucoma, Postoperative fibrosis in glaucoma Proliferative vitreoretinopathy (PVR), ·New blood vessels on the optic nerve disc, ·Corneal neovascularization, ·Retinal neovascularization, Vitreous neovascularization, Pannus, ·Pterygium, ·Vascular retinopathy, Diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (e.g., characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53) or proliferative diabetic retinopathy, e.g., in subjects not suffering from DME), and Diabetic retinopathy in subjects with diabetic macular edema (DME).

[0157] The scope of the present invention includes, for example, any of the methods described herein relating to nAMD, DR and / or DME, as well as methods related to intraocular neovascular diseases (e.g., ME-RVO) described herein.

[0158] The present invention provides a method for treating an intraocular neovascular disease (e.g., nAMD, DR, and / or DME) in a subject in need thereof by administering an initial loading dose (e.g., 2 mg or more, 4 mg or more, or preferably about 8 mg or more of a VEGF antagonist or inhibitor, e.g., a VEGF receptor fusion protein such as aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks, preferably every 4 weeks, preferably three initial loading doses), followed by successive additional doses about every 24 weeks, wherein the subject achieves, for example, by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks after initiation of treatment: an improvement in the Diabetic Retinopathy Severity Scale (DRSS), e.g., by at least 2 or 3 stages; -Improvement of best corrected visual acuity, Dry retina, · Obtaining best corrected visual acuity, Acquisition of best-corrected visual acuity of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 letters, or ≥ 5, ≥ 10, or ≥ 15 letters; ·BCVA of at least 69 letters, a decrease in central retinal thickness (CRT), for example by about 100, 125, 150, 175 or 200 micrometers, Absence of vascular leakage as measured by fluorescein angiography (FA), Improvement from pre-treatment baseline in the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) total score, The retina is free of fluid (total fluid, intraretinal fluid [IRF] and / or subretinal fluid [SRF]) in the fovea and central subfields; Preservation of fluid-free retina (total fluid in the fovea and central subfields, IRF and / or SRF) Macular edema loss, Fluid-free retina on spectral-domain optical coherence tomography (SD-OCT), and / or The present invention provides a method for achieving and / or maintaining non-departure from an HDq24 treatment regimen once initiated.

[0159] In embodiments of the invention, a subject undergoing HDq24 treatment for an intraocular neovascular disease (e.g., nAMD, DR and / or DME) described herein achieves one or more of the following: not undergo a dosing regimen change, for example, one in which the interval between doses (e.g., between tertiary doses) is shortened from a HDq24, HDq12-20, or HDq12, or HDq16, or HDq20 treatment regimen, once initiated, for example, by at least 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks; receiving 100% of all planned doses for, e.g., at least 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks; BVCA that is non-inferior to aflibercept administered intravitreally at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, then 2 mg approximately every 8 weeks or once every 2 months. an increase in BCVA (according to ETDRS letter score) of about 7, 8, or 9 letters by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks, e.g., a baseline BCVA of about 61, 62, and 63; -By 4 weeks after starting treatment, BCVA will improve by approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or by approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 8 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 12 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 16 weeks after starting treatment, patients will have an improvement in BCVA of approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 20 weeks after starting treatment, BCVA will improve by approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or by approximately 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 24 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 28 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 32 weeks after starting treatment, patients will have an improvement in BCVA of approximately 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 36 weeks after starting treatment, patients will have an improvement in BCVA of 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 40 weeks after starting treatment, BCVA will improve by approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 6 or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 44 weeks after starting treatment, patients will have an improvement in BCVA of approximately 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 48 weeks after initiation of treatment, an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen, e.g., a baseline BCVA of approximately 61, 62, or 63 letters (ETDRS or equivalent Snellen visual acuity); By 60 weeks after initiation of treatment, an improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen, e.g., a baseline BCVA of approximately 61, 62, or 63 letters (ETDRS or equivalent Snellen visual acuity); By 96 weeks after initiation of treatment, an improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 5, 6, or 7 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen, e.g., a baseline BCVA of approximately 60, 61, 62, or 63 letters (ETDRS or equivalent Snellen visual acuity); An improvement in BCVA of approximately 8 or 9 letters, or up to 40 letters (ETDRS or equivalent Snellen visual acuity) between weeks 48 and 60 if receiving an HDq12 or HDq24 regimen, for example, if baseline BCVA is approximately 63 or 64; or an improvement in BCVA of approximately 7 or 8 letters, or up to 40 letters (ETDRS or equivalent Snellen visual acuity) between weeks 48 and 60 if receiving an HDq16 or HDq24 regimen, for example, if baseline BCVA is approximately 61 or 62; An improvement in BCVA by approximately 8 weeks after initiation of treatment, which is maintained throughout the treatment regimen (e.g., within a range of approximately ±1 or ±2 ETDRS letters or equivalent Snellen visual acuity), for example, to at least 48 weeks; -Improvement in best corrected visual acuity (according to ETDRS letter score) (e.g., by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment), -Improvement in best corrected visual acuity (BVCA) by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after initiation of treatment; An increase in BCVA by ≥ 4 letters, ≥ 5 letters, ≥ 6 letters, ≥ 7 letters, ≥ 8 letters, ≥ 9 letters, or ≥ 10 letters (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after initiation of treatment), as measured, for example, by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or equivalent Snellen visual acuity; A change in BCVA score (according to ETDRS letter score) from the start of treatment of approximately 7, 8, or 9 between weeks 36 and 48, with a BCVA of approximately 60 or 70 at any time between weeks 36 and 48; Between weeks 36 and 48, if receiving HDq12 or HDq16 regimen, a change in BCVA score (according to ETDRS letter score) from the start of treatment of up to 38 letters, e.g., a baseline BCVA of approximately 27-79; A change in BCVA score (according to ETDRS letter score) from the start of treatment of approximately 7, 8, or 9 between weeks 48 and 60, with a BCVA of approximately 69, 70, 71, 72, or 73 at any time between weeks 48 and 60; - An improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) by 48 weeks after treatment initiation, for example, when the baseline BCVA is approximately ≤73 ETDRS letters and the patient is receiving an HDq12 or HDq24 regimen; - An improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) by, for example, 48 weeks after initiation of treatment, when the baseline BCVA is approximately ≥ 73 ETDRS letters and the patient is receiving an HDq12 or HDq24 regimen; - An improvement in BCVA of approximately 8 or 9 letters (ETDRS or equivalent Snellen visual acuity) by 48 weeks after treatment initiation, for example, when baseline BCVA is approximately ≤73 ETDRS letters, in patients receiving HDq16 or HDq24 regimens; - An improvement in BCVA of approximately 4 or 5 letters (ETDRS or equivalent Snellen visual acuity) by, for example, 48 weeks after initiation of treatment, when the baseline BCVA is approximately ≥ 73 ETDRS letters and the patient is receiving an HDq16 or HDq24 regimen; - An improvement in BCVA of approximately 7 or 8 letters (ETDRS or equivalent Snellen visual acuity) by, for example, 48 weeks after initiation of treatment, when receiving an HDq12 or HDq24 regimen and baseline CRT is < approximately 400 micrometers; - An improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) by 48 weeks after treatment initiation when receiving HDq12 or HDq24 regimens and baseline CRT is ≥ 400 micrometers; - An improvement in BCVA of approximately 5 or 6 letters (ETDRS or equivalent Snellen visual acuity) by, for example, 48 weeks after initiation of treatment, when receiving an HDq16 or HDq24 regimen and baseline CRT is < approximately 400 micrometers; - An improvement in BCVA of approximately 9 or 10 letters (ETDRS or equivalent Snellen visual acuity) by 48 weeks after initiation of treatment when receiving an HDq16 or HDq24 regimen and baseline CRT is ≥ approximately 400 micrometers; Have not lost 5, 10, or 15 letters (according to ETDRS letter score) by Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96; Obtain at least 5, 10, or 15 letters (according to ETDRS letter scores) by Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96; No loss of 5, 10, 15, or 69 or more letters of BCVA (e.g., 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks after initiation of treatment). Between 48 and 60 weeks, the BCVA score (according to the ETDRS letter score) is approximately 69, 70, 71, 72, or 73. BCVA (according to ETDRS letter score) of at least approximately 69 letters, for example, by week 48 or 60; By 4 weeks after starting treatment, the BCVA should be approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 66 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq24 or HDq16 regimen. By 8 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 67 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 12 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 16 weeks after starting treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 20 weeks after starting treatment, the BCVA should be approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 24 weeks after starting treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 67 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 28 weeks after starting treatment, the BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 32 weeks after starting treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 36 weeks after starting treatment, the BCVA should be approximately 71 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 68 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 40 weeks after starting treatment, the BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 69 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 44 weeks after starting treatment, the BCVA should be approximately 72 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 48 weeks after starting treatment, the BCVA should be approximately 73 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. By 96 weeks after starting treatment, the BCVA must be approximately 66, 67, 68, 69, or 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq12 or HDq24 regimen, or approximately 66, 67, 68, 69, or 70 letters (ETDRS or equivalent Snellen visual acuity) if receiving an HDq16 or HDq24 regimen. Between weeks 36 and 48, if receiving an HDq12 or HDq24 regimen, a BCVA of approximately 71, 72, 73, or 74 (ETDRS or equivalent Snellen visual acuity) when the baseline BCVA is approximately 57, 58, 59, 60, 61, 62, 63, or 64; or between weeks 36 and 48, if receiving an HDq16 or HDq24 regimen, a BCVA of approximately 69, 70, 71, 72, or 73 (ETDRS or equivalent Snellen visual acuity) when the baseline BCVA is approximately 55, 56, 57, 58, 59, 60, 61, or 62; If receiving an HDq12 or HDq24 regimen, for example, if the baseline BCVA is approximately 63 or 64, then between weeks 48 and 60 the BCVA will be approximately 69 or 70 letters, or up to 94 letters (ETDRS or equivalent Snellen visual acuity); or if receiving an HDq16 or HDq24 regimen, for example, if the baseline BCVA is approximately 61 or 62, then between weeks 48 and 60 the BCVA will be approximately 72 or 73 letters, or up to 89 letters (ETDRS or equivalent Snellen visual acuity); Achieving ≥5, ≥10, or ≥15 letters of BCVA (according to ETDRS letter score) (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks from the start of treatment), an improvement of ≥ 2 stages in the Diabetic Retinopathy Severity Scale (DRSS) (e.g., by week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 72, 84, 90, or 96 of treatment); an improvement of ≥3 stages in the Diabetic Retinopathy Severity Scale (DRSS) (e.g., by week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 72, 84, 90, or 96 of treatment); the retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the fovea or central subfield (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks after initiation of treatment) (as measured, for example, by optical coherence tomography (OCT)); Absence of intraretinal vascular leakage as measured by fluorescein angiography (FA) (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks after initiation of treatment), Maintenance of a fluid-free retina (total fluid in the fovea and central subfields, IRF, and / or SRF) (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks from the start of treatment), ETDRS grid (mm) at 48 or 60 weeks 2 ) the total area of ​​fluorescein leakage within the 2 or more, or up to approximately 57 or 68 mm 2 reducing (e.g., as measured by fluorescein angiography); ETDRS grid (mm) at 48 weeks 2 The total area of ​​fluorescein leakage within the 1000-kJ / kg / day (1000-kJ / kg / day) was approximately 13, 13.3, 13.9, or 14 mm2 when receiving HDq12 or HDq24 regimens. 2 or more (for example, up to approximately 52 mm 2 ) reduction (e.g., as measured by fluorescein angiography); ETDRS grid (mm) at 48 weeks 2 The total area of ​​fluorescein leakage within the 1000-kJ / kg / day (1000-kJ / kg / day) is approximately 7, 7.7, 8, 9, 9.4, or 10 mm2 when receiving HDq16 or HDq24 regimens. 2 or more (for example, up to approximately 55 mm 2 ) reduction (e.g., as measured by fluorescein angiography); - A fluid-free retina by spectral-domain optical coherence tomography (SD-OCT) at 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after the start of treatment; The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the fovea (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after initiation of treatment), Dry retina (e.g., by week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 after starting treatment), a fluid-free fovea (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks after initiation of treatment) (e.g., as measured by optical coherence tomography (OCT)); A change in central retinal thickness of approximately -118 or -118.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -124 or -125 or -124.9 or -125.5 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 4 weeks after starting treatment; A change in central retinal thickness of approximately -137 or -137.4 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -139 or -140 or -139.6 or -140.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 8 weeks after starting treatment; A change in central retinal thickness of approximately -150 or -150.1 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -152 or -153 or -152.7 or -153.4 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 12 weeks after starting treatment; A change in central retinal thickness of approximately -139 or -139.4 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -145 or -146 or -145.5 or -146.4 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 16 weeks after starting treatment; A change in central retinal thickness of approximately -117 or -117.1 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -112 or -113 or -112.5 or -113.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 20 weeks after starting treatment; A change in central retinal thickness of approximately -158 or -158.1 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -103 or -104 or -103.8 or -104.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 24 weeks after starting treatment; A change in central retinal thickness of approximately -146 or -147 or -146.7 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -162 or -162.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 28 weeks after starting treatment; A change in central retinal thickness of approximately -132 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -145, -146, or -145.8 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 32 weeks after starting treatment; A change in central retinal thickness of approximately -168 or -168.1 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -124 or -125 or -124.7 or -125.2 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 36 weeks after starting treatment; A change in central retinal thickness of approximately -163 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -122 or -123 or -122.5 or -123.1 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 40 weeks after starting treatment; A change in central retinal thickness of approximately -147, -148, or -147.4 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or approximately -164, -164.1, or -164.3 micrometers (± 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, by 44 weeks after starting treatment; a change in central retinal thickness of about -171 or -172 or -171.7, -172, -173, -174, -175, -176, or -176.77 micrometers (± 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or about -148 or -149 or -148.3 or -149.4 micrometers (± 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen, e.g., a baseline CRT of about 449, 450, 455, or 460 micrometers, by 48 weeks after initiation of treatment; Approximately -115, -116, -117, -118, -119, -120, -121, -122, -123, -124, -125, -126, -127, -128, -129, -130, -131, -132, -133, -134, -135, -136, -137, -138, -139, -140, -141, -142, -143, or -144 micrometers (± 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 micrometers) if receiving an HDq12 or HDq24 regimen, or if receiving an HDq16 or HDq24 regimen, by 96 weeks after starting treatment. a change in central retinal thickness of about -121, -122, -123, -124, -125, -126, -127, -128, -129, -130, -131, -132, -133, -134, -135, -136, -137, -138, -139, -140, -141, -142, -143, -144, -145, -146, -147, -148, -149, -150, -151, -152, -153 micrometers (± 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 micrometers) if the baseline CRT is about 350 or 370 micrometers; a change in central retinal thickness of about -181, -182, -181.95, -176, -176.24, or -177 (± 6, 10, 17, 18, or 19 micrometers) micrometers if receiving an HDq12 or HDq24 regimen (e.g., baseline CRT is about 460 micrometers), or about -166, -166.26, -167, or -167.18 micrometers (± 8, 9, 10, 17, 18, or 19 micrometers) if receiving an HDq16 or HDq24 regimen (e.g., baseline CRT is about 457 micrometers), by 60 weeks after initiation of treatment; A change in central retinal thickness of approximately -118, -119, or -118.3 micrometers between the start of treatment (week 0) and week 4 if receiving an HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -19, -20, or -19.1 micrometers between weeks 4 and 8 if receiving an HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -12, -13, or -12.7 micrometers between weeks 8 and 12 if receiving an HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -40 or -41 micrometers between weeks 20 and 24 if receiving HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -36, -37, or -36.1 micrometers between weeks 32 and 36 if receiving an HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -24, -25, or -24.3 micrometers between weeks 44 and 48 if receiving an HDq12 or HDq24 regimen, A change in central retinal thickness of approximately -124, -125, or -124.9 micrometers between the start of treatment (week 0) and week 4 if receiving an HDq16 or HDq24 regimen, A change in central retinal thickness of approximately -14, -15, or -14.7 micrometers between weeks 4 and 8 if receiving an HDq16 or HDq24 regimen, A change in central retinal thickness of approximately -13, -14, or -13.1 micrometers between weeks 8 and 12 if receiving an HDq16 or HDq24 regimen, A change in central retinal thickness of approximately -58, -59, or -58.5 micrometers between weeks 24 and 28 if receiving an HDq16 or HDq24 regimen, A change in central retinal thickness of approximately -41, -42, or -41.6 micrometers between weeks 40 and 44 if receiving an HDq16 or HDq24 regimen, a reduction in CRT by week 4, 5, 6, 7, or 8 after initiation of treatment, which is maintained thereafter throughout the treatment regimen, for example, to at least week 48 (e.g., within about ±17, ±18, or ±19 micrometers); a decrease in central retinal thickness (CRT) of, for example, about 100, 125, 150, 175, or 200 micrometers (by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after the start of treatment); a decrease in CRT (e.g., as measured by optical coherence tomography (OCT)) of about 148 to 182 micrometers (e.g., 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183) by week 48 or 60, with a baseline CRT of about 449, 450, 455, or 460 micrometers; a decrease in central retinal thickness (CRT), e.g., by at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 micrometers (e.g., by 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 weeks after initiation of treatment); that the ocular (e.g., intraocular pressure) and non-ocular safety (e.g., hypertensive events or APTC events) or mortality rates in subjects with intraocular neovascular diseases, such as DR or DME, are similar to those of aflibercept administered intravitreally at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, followed by 2 mg once every 8 weeks or once every 2 months; about 0.1667 days after the first administration, the plasma free aflibercept concentration is about 0.149 (±0.249) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 0.3333 days after the first administration, the plasma free aflibercept concentration is about 0.205 (±0.250) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 1 day after the first administration, the plasma free aflibercept concentration is about 0.266 (±0.211) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 2 days after the first administration, the plasma free aflibercept concentration is about 0.218 (±0.145) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 4 days after the first administration, the plasma free aflibercept concentration is about 0.140 (±0.0741) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 7 days after the first administration, the plasma free aflibercept concentration is about 0.0767 (±0.0436) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 14 days after the first administration, the plasma free aflibercept concentration is about 0.0309 (±0.0241) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 21 days after the first administration, the plasma free aflibercept concentration is about 0.0171 (±0.0171) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 28 days after the first administration, the plasma free aflibercept concentration is about 0.00730 (±0.0113) mg / L, e.g., the plasma free aflibercept concentration at baseline is undetectable, e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 0.1667 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.00698 (±0.0276) mg / L, and the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 0.3333 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.00731 (±0.0279) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 1 day after the first administration, the plasma adjusted conjugated aflibercept concentration is about 0.0678 (±0.0486) mg / L, e.g., the baseline adjusted conjugated aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 2 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.138 (±0.0618) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 4 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.259 (±0.126) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 7 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.346 (±0.151) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 14 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.374 (±0.110) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 21 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.343 (±0.128) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; about 28 days after the first administration, the plasma adjusted bound aflibercept concentration is about 0.269 (±0.149) mg / L, e.g., the baseline adjusted bound aflibercept concentration is about 0.00583 mg / L (±0.0280), e.g., the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks; - the peak plasma concentration of free aflibercept is reached at about 0.965 (e.g., about 1) days after the first administration; - Reaching a peak plasma concentration of free aflibercept of approximately 0.310 mg / l (±0.263); Individual free aflibercept plasma concentrations (C max ) is about 0 to about 1.08 mg / L (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 or 1.1 mg / L), The maximum plasma free aflibercept concentration (mg / L) per mg aflibercept dose is approximately 0.0388 (±0.00328) mg / L / mg. - Peak plasma concentrations of adjusted conjugated aflibercept are reached approximately 14 days after the first dose; Achieving a peak plasma concentration of approximately 0.387 mg / l (±0.135) of adjusted bound aflibercept; - adjusted conjugated aflibercept plasma concentration of approximately 0.137 to approximately 0.774 mg / L (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 mg / L); - the peak plasma concentration (mg / L) of adjusted conjugated aflibercept per mg dose of aflibercept is approximately 0.0483 (±0.0168) mg / L / mg; - Absence of anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment an improvement from pre-treatment baseline (e.g., by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment) in the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) total score, e.g., by about 4, 5, or 6 if receiving an HDq12 regimen or an HDq24 regimen, or by about 2, 3, or 4 if receiving an HDq16 regimen or an HDq24 regimen, e.g., a baseline score of about 76 or 77; Resolution of macular edema (e.g., by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks after initiation of treatment), and / or In subjects with DR or DME, the efficacy and / or safety is similar to that of aflibercept administered intravitreally at 2 mg approximately every 4 weeks for the first 5 injections, then 2 mg once every 8 weeks or once every 2 months, where efficacy is measured, for example, as an increase in BCVA and / or a decrease in central retinal thickness, and safety is measured, for example, as the incidence of adverse events (treatment-emergent adverse events occurring any time within 30 days of any injection), such as increased blood pressure, increased intraocular pressure, visual disturbances, vitreous floaters, vitreous detachment, iris neovascularization, and / or vitreous hemorrhage.

[0160] Thus, the present invention provides: A method for achieving non-inferior BVCA compared to aflibercept administered intravitreally at 2 mg every 4 weeks for the first 3, 4, or 5 injections, then approximately once every 8 weeks or once every 2 months, in a subject in need thereof with intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial administration of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration (HD24 regimen). A method for achieving an increase in BCVA (according to ETDRS letter score) of about 7, 8, or 9 letters by Week 60 in a subject with intraocular neovascular disease (preferably DR and / or DME), where the increase is from a baseline BCVA of about 61, 62, or 63, comprising administering to the subject's eye a single initial administration of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, where each secondary administration is administered about 2-4 weeks after the immediately preceding administration, and where each tertiary administration is administered about 24 weeks after the immediately preceding administration (HDq24 regimen). A method for achieving a BCVA (according to ETDRS letter score) of at least 69 letters by Week 48 or 60 in a subject in need thereof having an intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial dose of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary doses of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary doses of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose (HDq24 regimen). A method for achieving a BCVA with no loss of 5, 10, 15, or 69 letters after 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks of treatment in a subject in need thereof having an intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial dose of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary doses of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary doses of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose (HDq24 regimen). A method for achieving an improvement in best-corrected visual acuity (according to ETDRS letter score) by 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks from the start of treatment in a subject in need thereof having an intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial dose of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary doses of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary doses of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary dose is administered about 2-4 weeks after the immediately preceding dose, and each tertiary dose is administered about 24 weeks after the immediately preceding dose (HDq24 regimen). A method for achieving an improvement in best corrected visual acuity (BVCA) by 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 32 weeks, 36 weeks, 40 weeks, 44 weeks, 48 ​​weeks, or 60 weeks after initiation of treatment in a subject in need thereof having an intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial administration of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration and each tertiary administration is administered about 24 weeks after the immediately preceding administration (HDq24 regimen). A method for achieving a BCVA score (according to ETDRS letter scores) of about 69, 70, 71, 72, or 73 between 48 and 60 weeks from the start of treatment in a subject in need thereof having an intraocular neovascular disease (preferably DR and / or DME), comprising administering to the subject's eye a single initial administration of at least about 8 mg of a VEGF receptor fusion protein, followed by one or more secondary administrations of at least about 8 mg of the VEGF receptor fusion protein, followed by one or more tertiary administrations of at least about 8 mg of the VEGF receptor fusion protein, wherein each secondary administration is administered about 2-4 weeks after the immediately preceding administration, and each tertiary administration is administered about 24 weeks after the immediately preceding administration (HDq24 regimen). A method for achieving a change in BCVA score (according to ETDRS letter score) from the start of treatment of about 7, 8, or 9 between 36 and 48 weeks in a subject in n...

Claims

1. - In subjects requiring treatment or prevention of intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy (DR), and / or diabetic macular edema (DME), for the purpose of providing said treatment or prevention, - In subjects with nAMD, DR, and / or DME who require improvement of their best corrected visual acuity, for the purpose of performing such improvement, - A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method for promoting retinal dryness in subjects having intraocular neovascular disease, nAMD, DR and / or DME, A pharmaceutical composition comprising administering approximately 8 mg or more of a VEGF receptor fusion protein to the target eye once every 24 weeks.

2. - A pharmaceutical composition according to claim 1 for use in a method for treating or preventing intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy (DR), and / or diabetic macular edema (DME) in subjects requiring such treatment or prevention, The method applies to the target eye, A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by One or more secondary doses of approximately 8 mg or more of VEGF receptor fusion protein, followed by This includes administering approximately 8 mg or more of VEGF receptor fusion protein as a third dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. The pharmaceutical composition according to claim 1, wherein each third dose is administered approximately 24 weeks after the immediately preceding dose.

3. Intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy ( A pharmaceutical composition according to claim 1 for use in a method for treating or preventing DR) and / or diabetic macular edema (DME) in a subject requiring treatment or prevention, The method applies to the target eye, A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by The second dose is approximately 8 mg or more of VEGF receptor fusion protein, followed by This includes administering approximately 8 mg or more of VEGF receptor fusion protein as a third dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. The pharmaceutical composition according to claim 1, wherein each third dose is administered approximately 24 weeks after the immediately preceding dose.

4. A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method for treating or preventing intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy and / or diabetic macular edema in subjects requiring such treatment or prevention, wherein the method comprises administering a formulation containing approximately 114.3 mg / ml of VEGF receptor fusion protein to the eye of the subject in three doses of approximately 8 mg of VEGF receptor fusion protein, once every four weeks, and then administering one or more doses of VEGF receptor fusion protein after the three doses, with intervals extended up to a maximum of 24 weeks.

5. A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method for slowing the clearance of free aflibercept from the ocular compartment after intravitreous injection, compared to the rate of clearance of aflibercept from the ocular compartment after intravitreous injection of 2 mg or ≤4 mg of aflibercept, wherein the VEGF receptor fusion protein is aflibercept, and the method is To the eyes of the target that needs it, Aflibercept, administered as a single initial dose of approximately 8 mg or more, followed by Aflibercept, administered as a secondary dose of approximately 8 mg or more, followed by This includes intravitreal injection of approximately 8 mg or more of aflibercept as a third-line dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. A pharmaceutical composition in which each third dose is administered approximately 24 weeks after the immediately preceding dose.

6. The pharmaceutical composition according to claim 1, wherein the clearance of free aflibercept from the ocular compartment is about 34% slower than the clearance from the ocular compartment after intravitreal injection of 2 mg or ≤4 mg of aflibercept.

7. The pharmaceutical composition according to claim 1, wherein the clearance of free aflibercept from the ocular compartment is approximately 0.37 to 0.46 mL / day or 0.41 mL / day after intravitreal injection of ≥ 8 mg of aflibercept.

8. A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method to increase the duration of efficacy and / or the time it takes for the amount of free aflibercept to reach the lower limit of quantification (LLOQ) in the target ocular compartment after intravitreal injection of aflibercept, compared to the time it takes for the amount of free aflibercept to reach the lower limit of quantification (LLOQ) in the target ocular compartment after intravitreal injection of aflibercept, wherein the VEGF receptor fusion protein is aflibercept, and the method is used to deliver to the eye of the target as required. Aflibercept, administered as a single initial dose of approximately 8 mg or more, followed by Aflibercept, administered as a secondary dose of approximately 8 mg or more, followed by This includes intravitreal injection of approximately 8 mg or more of aflibercept as a third-line dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. A pharmaceutical composition in which each third dose is administered approximately 24 weeks after the immediately preceding dose.

9. The pharmaceutical composition according to claim 8, wherein the duration of efficacy and / or the time it takes for the amount of free aflibercept to reach the lower limit of quantification (LLOQ) in the target ocular compartment after intravitreal injection of aflibercept is increased by about 5 or 6 weeks compared to the time it takes for the amount of free aflibercept in the target ocular compartment to reach LLOQ after intravitreal injection of about 2 mg or ≤4 mg of aflibercept.

10. The pharmaceutical composition according to claim 8, wherein the time it takes for the amount of free aflibercept in the target ocular compartment to reach the lower limit of quantification (LLOQ) after intravitreal injection of aflibercept is increased by approximately 1, 2, 1.2, or 1.3 weeks or more compared to the time it takes for the amount of free aflibercept in the target ocular compartment to reach LLOQ after intravitreal injection of approximately 2 mg or ≤4 mg of aflibercept.

11. The pharmaceutical composition according to claim 8, wherein the time it takes for the amount of free aflibercept in the target ocular compartment to reach the lower limit of quantification (LLOQ) after intravitreal injection of ≥8 mg of aflibercept is approximately 15 weeks.

12. The pharmaceutical composition according to claim 8, wherein the time it takes for the amount of free aflibercept in the target ocular compartment to reach the lower limit of quantification (LLOQ) after intravitreal injection of ≥8 mg of aflibercept is longer than approximately 8, 8.7, 8.71, 9, 9.2, 9.21, or 10 weeks.

13. A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method to increase the time it takes for free aflibercept in the plasma of a subject to reach the lower limit of quantification (LLOQ) after intravitreal injection of aflibercept, compared to the time it takes for free aflibercept in the plasma of a subject to reach LLOQ after intravitreal injection of approximately 2 mg or ≤4 mg of aflibercept, wherein the VEGF receptor fusion protein is aflibercept, and the method is used in the eye of a subject as required. Aflibercept, administered as a single initial dose of approximately 8 mg or more, followed by Aflibercept, administered as a secondary dose of approximately 8 mg or more, followed by This includes intravitreal injection of approximately 8 mg or more of aflibercept as a third-line dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. A pharmaceutical composition in which each third dose is administered approximately 24 weeks after the immediately preceding dose.

14. The pharmaceutical composition according to claim 13, wherein the LLOQ of free aflibercept measured in plasma is approximately 0.0156 mg / L.

15. The pharmaceutical composition according to claim 14, wherein the time it takes for free aflibercept in the target plasma to reach the lower limit of quantification (LLOQ) after intravitreal injection of aflibercept is increased by about two weeks compared to the time it takes for free aflibercept in the target plasma to reach LLOQ after intravitreal injection of about 2 mg of aflibercept.

16. The pharmaceutical composition according to claim 15, wherein the time it takes for free aflibercept in the target plasma to reach the lower limit of quantification (LLOQ) after intravitreal injection of ≥8 mg of aflibercept is approximately 3, 3.5, 3.8, or 4 weeks.

17. The pharmaceutical composition according to claim 16, wherein the time it takes for free aflibercept in the target plasma to reach the lower limit of quantification (LLOQ) after intravitreal injection of ≥8 mg of aflibercept is longer than approximately 1.5 or 1.6 weeks.

18. The pharmaceutical composition according to claim 13, wherein ≤4 mg is approximately 2 mg or 2 to 4 mg.

19. The pharmaceutical composition according to claim 13, wherein the subject suffers from an intraocular neovascular disease, age-related macular degeneration with neovascularization, diabetic retinopathy, and / or diabetic macular edema.

20. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation containing a histidine buffer.

21. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation containing arginine.

22. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation having a pH of approximately 5.

8.

23. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation containing a sugar or a polyol.

24. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation containing sucrose.

25. The pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical formulation, and the aflibercept has high molecular weight molecular species of less than about 3.5% immediately after manufacture and purification, and / or high molecular weight molecular species of less than about 6% after storage at about 2 to 8°C for about 24 months.

26. A pharmaceutical composition according to any one of claims 1 to 19, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical preparation comprising an aqueous pharmaceutical preparation, the aqueous pharmaceutical preparation comprising at least about 100 mg / ml of a VEGF receptor fusion protein containing two polypeptides each comprising the second immunoglobin-like (Ig) domain of VEGFR1, the third Ig domain of VEGFR2, and a polymer-forming component, about 10 to 100 mM of L-arginine, sucrose, a histidine-based buffer, and a surfactant, the aqueous pharmaceutical preparation having a pH of about 5.0 to about 6.8, and the VEGF receptor fusion protein having less than about 3.5% of high molecular weight molecular species immediately after production and purification, and / or about 6% or less of high molecular weight molecular species after storage at about 2 to 8°C for about 24 months.

27. - In subjects requiring treatment or prevention of intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy (DR), and / or diabetic macular edema (DME), for the purpose of providing said treatment or prevention, - In subjects with nAMD, DR, and / or DME who require improvement of their best corrected visual acuity, for the purpose of performing such improvement, - A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method for promoting retinal dryness in subjects having intraocular neovascular disease, nAMD, DR and / or DME, The method applies to the eye of the subject, A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by One or more secondary doses of approximately 8 mg or more of VEGF receptor fusion protein, followed by This includes administering approximately 8 mg or more of VEGF receptor fusion protein as a third dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. Each third dose is administered approximately 8 to 23 weeks after the previous dose, and the subjects are as follows:

1. Loss of 5 letters in BCVA, 2. CRT is <300 μm or <320 μm, 3. The OCT shows no liquid in the central subfield, and / or 4. If there is no new onset of foveal hemorrhage or foveal neovascularization, A pharmaceutical composition comprising extending the interval between third doses by one week or more.

28. The pharmaceutical composition according to claim 27, wherein the interval between the third doses has been extended at least once so far.

29. The interval between the aforementioned third doses has occurred at least once so far. From 8 weeks to 12, 16, or 20 weeks, 12 weeks to 16 or 20 weeks, and / or The pharmaceutical composition according to claim 27, extended from 16 weeks to 20 weeks.

30. The pharmaceutical composition according to claim 27, wherein extending the interval between the third doses by one week or more means extending the interval between the third doses by one or more increments of four weeks.

31. The pharmaceutical composition according to claim 27, wherein the 8 to 23 weeks is 8, 12, 16, or 20 weeks.

32. The pharmaceutical composition according to claim 31, wherein the aforementioned 8 to 23 weeks is 20 weeks.

33. The interval between the third doses is such that the subject is 1. Loss of the first 5 letters of BCVA, and 2. The pharmaceutical composition according to claim 27, which is extended when the CRT exhibits a width of <300 μm or <320 μm.

34. The pharmaceutical composition according to claim 33, wherein the subject has DR and / or DME.

35. The interval between the third doses is such that the subject is 1. Loss of the first 5 letters of BCVA, and 2. There is no liquid in the central subfield, and 3. The pharmaceutical composition according to claim 27, which is extended in cases where there is no newly occurring foveal hemorrhage or foveal neovascularization.

36. The pharmaceutical composition according to claim 35, wherein the subject has nAMD.

37. - Loss of <5 letters in BCVA refers to the loss of <5 letters in BCVA starting from week 12 of treatment. • CRTs with a diameter of <300 μm or <320 μm are derived from SD-OCT. • The CRT with a diameter of <300 μm is from Cirrus SD-OCT. • CRTs <320 μm are from Spectralis SD-OCT, and / or The pharmaceutical composition according to claim 27, wherein the absence of liquid in the central subfield is on the OCT.

38. - In subjects requiring treatment or prevention of intraocular neovascular disease, age-related macular degeneration with neovascularization (nAMD), diabetic retinopathy (DR), and / or diabetic macular edema (DME), for the purpose of providing said treatment or prevention, - In subjects with nAMD, DR, and / or DME who require improvement of their best corrected visual acuity, for the purpose of performing such improvement, - A pharmaceutical composition comprising a VEGF receptor fusion protein for use in a method for promoting retinal dryness in subjects having intraocular neovascular disease, nAMD, DR and / or DME, The method applies to the eye of the subject, A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by One or more secondary doses of approximately 8 mg or more of VEGF receptor fusion protein, followed by This includes administering approximately 8 mg or more of VEGF receptor fusion protein as a third dose, Each secondary dose is administered approximately 2 to 4 weeks after the previous dose. Each third dose is administered approximately 24 weeks after the previous dose, and the subject is:

1. Loss of >5 or >10 characters in BCVA, 2. An increase of >25 μm or >50 μm in central retinal thickness (CRT), 3. Novel foveal hemorrhage, 4. Novel foveal neovascularization, and / or 5. If DME persists or worsens, A pharmaceutical composition comprising shortening the interval between third doses by one week or more.

39. The pharmaceutical composition according to claim 38, wherein shortening the interval between the third doses by one week or more means shortening the interval between the third doses by one or more increments of four weeks.

40. The interval between the third doses is such that the subject is 1. In association with the persistence or worsening of DME, BCVA is >10 letter loss, and 2. The pharmaceutical composition according to claim 38, which is extended when it exhibits an increase of >50 μm in CRT.

41. The pharmaceutical composition according to claim 40, wherein the subject has DR and / or DME.

42. The interval between the third doses is such that the subject is 1. Loss of >5 letters in BCVA, and 2. The pharmaceutical composition according to claim 38, which is extended when the central retinal thickness (CRT) increases by >25 μm, or when new foveal hemorrhage or new foveal neovascularization is present.

43. The pharmaceutical composition according to claim 42, wherein the subject has nAMD.

44. - Loss of >10 letters in BCVA in association with the persistence or worsening of DME means that the loss of >10 letters in BCVA starting from week 12 of treatment in association with the persistence or worsening of DME. • The increase in CRT >50 μm occurred from week 12 of treatment initiation. The loss of the >5 letters in BCVA occurred from the 12th week after the start of treatment. The pharmaceutical composition according to claim 38, wherein an increase of >25 μm in central retinal thickness (CRT), or a new foveal hemorrhage, or a new foveal neovascularization occurs from the 12th week after the start of treatment.

45. By the 96th week from the start of treatment, the aforementioned subjects - Improvement of BCVA of approximately 6, 7, 8, or 9 characters. - Improvement of BCVA with 39, 40, 41, 42, 43, 44, 45, 46, or 47 characters. - BCVA must not lose 5 or more characters, 10 or more characters, or 15 or more characters. - BCVA must be approximately 69, 70, 71, 72, or 73 characters long. - BCVA must have 5 or more characters, 10 or more characters, or 15 or more characters. - Improvement of DRSS by two or more levels. - A decrease in CRT of approximately 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, or 185 micrometers. CRTs of approximately 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, or 304 micrometers, No IRF, - No SRF, - No IRF, and no SRF, - No significant change in intraocular pressure. - No significant change in systolic blood pressure, and / or A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, which exhibits one or more of the following: - No significant change in diastolic blood pressure.

46. The pharmaceutical composition according to claim 45, wherein the subject has DR and / or DME.

47. By the 96th week from the start of treatment, the aforementioned subjects - Improvement of BCVA of approximately 5, 6, or 7 characters. - Improvement of BCVA with 43, 44, 45, 46, 47, or 48 characters. ・BCVA must be 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, or 94 characters long. - BCVA must not lose 5 or more characters, 10 or more characters, or 15 or more characters. - BCVA must have 5 or more characters, 10 or more characters, or 15 or more characters. - Improvement of DRSS by two or more levels. • Approximately 12; 13; 14; 15; 16; 17; 18; 19; 20; 21; 22; 23; 24; 25; 26; 27; 28; 29; 30; 31; 32; 33; 34; 35; 36; 37; 38; 39; 40; 41; 42; 43; 44; 45; 46; 47; 48; 49; 50; 51; 52; 53; 54; 55; 56; 57; 58; 59; 60; 61;62;63;64;65;66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83;84;85;86;87;88;89;90;91;92;93;94;95;96;97;98;99;100;101;102;103;104;105;106;107;1 08; 109; 110; 111; 112; 113; 114; 115; 116; 117; 118; 119; 120; 121; 122; 123; 124; 125; 126; 127; 128; 129; 130; 131; 132; 133; 134; 135; 136; 137; 138; 139; 140; 141; 142; 143; 144; 145 ;146;147;148;149;150;151;152;153;154;155;156;157;158;159;160;161;162;163;164;165;166;167;168;169;170;171;172;173;174;175;176;177;178;179;180;181;182;1 83;184;185;186;187;188;189;190;191;192;193;194;195;196;197;198;199;200;201;202;203;204;205;206;207;208;209;210;211;212;213;214;215;216;217;218;219;220;221;222;223;224;225;226;227;228;229;230;231;232;233;234;235;236;237;238;239;240;2 CRT reduction of 41; 242; 243; 244; 245; 246; 247; 248; 249; 250; 251; 252; 253; 254; 255; 256; 257; 258; 259; 260; 261; 262; 263; 264; 265; 266; 267; 268; 269; 270; 271; 272; 273; 274; 275; 276; 277; 278; 279; 280; 281; 282; 283; 284; 285; 286; 287; 288; 289; 290; 291; 292; 293, or 294 micrometers. - CRT is approximately 156, 157; 158; 159; 160; 161; 162; 163; 164; 165; 166; 167; 168; 169; 170; 171; 172; 173; 174; 175; 176; 177; 178; 179; 180; 181; 182; 183; 184; 185; 186; 187; 1 88; 189; 190; 191; 192; 193; 194; 195; 196; 197; 198; 199; 200; 201; 202; 203; 204; 205; 206; 207; 208; 209; 210; 211; 212; 213; 214; 215; 216; 217; 218; 219; 220; 221; 22 2;223;224;225;226;227;228;229;230;231;232;233;234;235;236;237;238;239;240;241;242;243;244;245;246;247;248;249;250;251;252;253;254;255;25 6;257;258;259;260;261;262;263;264;265;266;267;268;269;270;271;272;273;274;275;276;277;278;279;280;281;282;283;284;285, or 286 micrometers. Approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mm 2 The decrease in choroidal neovascularization size, - Choroidal neovascularization size is approximately 0, 1, 2, or 3 mm. 2 Being - No leakage should be observed on fluorescein angiography. - Total lesion size is approximately 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 mm 2 Being Approximately 0, 0.1, 0.2, or 0.3 mm 2 Reduction in total lesion size, - The total score of the NEI-VFQ-25 is approximately 80. - The change in the total NEI-VFQ-25 score from baseline is approximately 2 or 3. No IRF, - No SRF, - No IRF, and no SRF, - No significant change in intraocular pressure. - No significant change in systolic blood pressure, and / or A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, which exhibits one or more of the following: - No significant change in diastolic blood pressure.

48. The pharmaceutical composition according to claim 47, wherein the subject has nAMD.

49. Eye infection, or periorbital infection, Active intraocular inflammation, and / or Subjects who have one or more of the following hypersensitivity conditions: Claims 1-19 and 27 are excluded from the administration of VEGF receptor fusion proteins to the eye. A pharmaceutical composition according to any one of items 44 to 44.

50. The aforementioned method targets, Eye infection, or periorbital infection, Active intraocular inflammation, and / or Steps for evaluating hypersensitivity, The pharmaceutical composition according to claim 49, further comprising the step of excluding any one or more of the subjects from the administration if any of the subjects are found.

51. The pharmaceutical composition according to claim 49, further comprising monitoring the subject during the treatment or prevention of conjunctival hemorrhage, cataract, vitreous detachment, vitreous floaters, corneal epithelial defects, and / or elevated intraocular pressure.

52. The above method involves packaging together before each dose. A single-dose glass vial with a protective plastic cap and stopper contains an aqueous formulation containing 8 mg of VEGF receptor fusion protein in approximately 70 microliters. - A single 18-gauge x 1.5-inch, 5-micron filter needle including the tip and beveled end, - One 30 gauge x 0.5 inch hypodermic needle, To provide a 1-mL Luer lock syringe having a graduated line marking a volume of 70 microliters, Next, (1) Visually inspect the aqueous formulation in the vial, and if particles, turbidity, or discoloration are observed, then use another vial of the aqueous formulation containing the VEGF receptor fusion protein. (2) Remove the protective plastic cap from the vial, (3) Clean the top of the vial with an alcohol wipe, and then, By using aseptic techniques, (4) Remove the 18 gauge x 1.5 inch, 5 micron filter needle and the 1 mL syringe from their packaging, (5) Attaching the filter needle to the syringe by twisting it onto the tip of the Luer lock syringe, (6) Push the filter needle into the center of the vial stopper until the needle is fully inserted into the vial and the tip of the needle contacts the bottom or bottom edge of the vial, (7) With the vial upright and slightly tilted, while ensuring that the angled end of the filter needle is immersed in the liquid, the contents of the VEGF receptor fusion protein vial are drawn into the syringe, (8) During withdrawal, tilt the vial so that the angled end of the filter needle remains immersed in the formulation, (9) When emptying the vial to completely empty the filter needle, fully retract the plunger rod, (10) Remove the filter needle from the syringe and discard the filter needle, (11) Remove the 30 gauge x 0.5 inch injection needle from its packaging and attach the injection needle to the syringe by screwing it firmly onto the tip of the Luer lock syringe, (12) Hold the syringe with the needle pointing upwards and check for air bubbles in the syringe; if air bubbles are present, gently flick the syringe with your finger until the bubbles rise to the top. (13) The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, comprising: slowly pushing the plunger so that the tip of the plunger aligns with a mark on the syringe indicating 70 microliters.

53. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the injection of the VEGF receptor fusion protein is performed under controlled sterile conditions, which include surgical hand disinfection, as well as the use of sterile gloves, sterile drapes, and sterile eyelid retractors (or equivalents), and an anesthetic and a topical broad-spectrum antimicrobial eye drop is administered prior to the injection.

54. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein comprises amino acids 27 to 457 of the amino acid sequence described in SEQ ID NO:

2.

55. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is selected from the group consisting of aflibercept and convert.

56. The aforementioned VEGF receptor fusion protein, (i) comprising two polypeptides: (1) VEGFR1 component containing amino acids 27-129 of SEQ ID NO: 2, (2) VEGFR2 component containing amino acids 130-231 of SEQ ID NO: 2, and (3) a polymer-forming component containing amino acids 232-457 of SEQ ID NO:

2. (ii) comprising the second immunoglobin-like (Ig) domain of VEGFR1, the third Ig domain of VEGFR2, and two polypeptides containing a polymerizing component, (iii) comprising two polypeptides containing the second immunoglobin-like (Ig) domain of VEGFR1, the third Ig domain of VEGFR2, the fourth Ig domain of VEGFR2 and a polymerizing component, or (iv) The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, comprising two VEGFR1R2-FcΔC1(a) polypeptides encoded by the nucleic acid sequence of Sequence ID No.

1.

57. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein comprises a second immunoglobin-like (Ig) domain of VEGFR1, a third Ig domain of VEGFR2, and two polypeptides comprising a polymerizing component.

58. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation selected from the group consisting of A to KKKK.

59. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation containing approximately 114.3 mg / ml of VEGF receptor fusion protein.

60. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the method comprises administering the VEGF receptor fusion protein to both eyes of the subject.

61. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is administered from a pre-filled syringe.

62. The pharmaceutical composition according to claim 61, wherein the pre-filled syringe is made of glass or plastic and / or is sterilized.

63. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is injected intravitreously using a 30-gauge x 0.5-inch sterile injection needle.

64. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the subject has previously received one or more doses of 2 mg of VEGF receptor fusion protein.

65. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein one or more further doses of VEGF receptor fusion protein are administered.

66. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 2 mg of VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation containing approximately 40 mg / ml of VEGF receptor fusion protein.

67. 2 mg of VEGF receptor fusion protein, A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, comprising 40 mg / ml of VEGF receptor fusion protein, 10 mM sodium phosphate, 40 mM NaCl, 0.03% polysorbate 20, and 5% sucrose, and having a pH of 6.2, in an aqueous pharmaceutical formulation.

68. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 8 mg of VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation containing a sugar or a polyol.

69. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 8 mg of VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation containing sucrose.

70. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 8 mg of VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation having a pH of approximately 5.

8.

71. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 8 mg of VEGF receptor fusion protein is present in an aqueous pharmaceutical formulation containing approximately 103 to 126 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine.

72. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein 8 mg of VEGF receptor fusion protein is an aqueous pharmaceutical preparation containing approximately 114.3 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine.

73. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein ≥ 8 mg of aflibercept is present in an aqueous pharmaceutical formulation, and the aflibercept has high molecular weight molecular species of less than about 3.5% immediately after manufacture and purification, and / or high molecular weight molecular species of less than about 6% after storage at about 2 to 8°C for about 24 months.

74. ≥8 mg of VEGF receptor fusion protein, At least approximately 100 mg / ml of VEGF receptor fusion protein, Approximately 10-100 mM L-arginine, Sucrose and, Histidine-based buffer and It is contained in an aqueous pharmaceutical formulation containing a surfactant, The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the aqueous pharmaceutical preparation has a pH of about 5.0 to about 6.8, and the VEGF receptor fusion protein has high molecular weight molecular species of less than about 3.5% immediately after production and purification, and / or high molecular weight molecular species of 6% or less after storage at about 2 to 8°C for about 24 months.

75. Approximately 8 mg of VEGF receptor fusion protein is present in the aqueous pharmaceutical formulation, and the aqueous pharmaceutical formulation is • Contains >100 mg / ml of VEGF receptor fusion protein, histidine-based buffer, and L-arginine, - Contains 140 mg / ml aflibercept, 20 mM histidine buffer, 5% sucrose, 0.03% polysorbate 20, 10 mM L-arginine, pH 5.8, - Contains 150 + 15 mg / ml aflibercept, 10 mM phosphate buffer, 8 ± 0.8% (w / v) sucrose, 0.02–0.04% (w / v) polysorbate 20 and 50 mM L-arginine, pH 5.9–6.5, - Contains 103-126 mg / ml of aflibercept, 10 ± 1 mM histidine-based buffer, 5 ± 0.5% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20, and 50 ± 5 mM L-arginine, pH 5.5-6.1, - Contains 140 mg / ml aflibercept, 10 mM histidine buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8, - Contains 114.3 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine, pH 5.8, • Contains aflibercept > 100 mg / ml, histidine-based buffer, and L-arginine, - Aflibercept in a concentration of >100 mg / ml with a pH of approximately 5.8, wherein the aqueous pharmaceutical formulation contains >100 mg / ml of aflibercept, which forms <3% HMW aggregates after incubation at 5°C for 2 months. - Contains approximately 114.3 mg / mL of aflibercept, 10 mM to 50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, and pH 5.8, or A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, comprising approximately 114.3 mg / mL of aflibercept; 10 mM His / His-HCl buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, and pH 5.

8.

76. 8 mg of VEGF receptor fusion protein in approximately 100 μl or less, approximately 75 μl or less, approximately 70 μl or less, or approximately 50 μl, 51 μl, 52 μl, 53 μl, 54 μl, 55 μl, 56 μl, 57 μl, 58 μl, 59 μl, 60 μl, 61 μl, 62 μl, 63 μl, 64 μl, 65 μl, 66 μl, 67 μl, 68 μl, 69 μl, 70 μl, 71 μl, 72 μl, 73 μl, 74 μl, A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, administered in a volume of 75 μl, 76 μl, 77 μl, 78 μl, 79 μl, 80 μl, 81 μl, 82 μl, 83 μl, 84 μl, 85 μl, 86 μl, 87 μl, 88 μl, 89 μl, 90 μl, 91 μl, 92 μl, 93 μl, 94 μl, 95 μl, 96 μl, 97 μl, 98 μl, 99 μl, or 100 μl.

77. The pharmaceutical composition according to claim 76, wherein the VEGF receptor fusion protein is administered in a volume of approximately 70 ± 4 or 5 microliters.

78. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the interval between administrations is adjusted (increased / maintained / decreased) based on visual outcomes and / or anatomical outcomes.

79. The above method may, as needed, be performed as needed (pro renata) (PRN), capped A pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, further comprising one or more periods of administration of PRN or Treat and Extended (T&E).

80. The pharmaceutical composition according to any one of claims 1 to 19 and 27 to 44, wherein the VEGF receptor fusion protein is aflibercept.

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