Prolonged high dose VEGF antagonist regimen for treatment of angiogenic eye diseases

Abstract

The present invention relates to a regimen for the treatment of angiogenic eye diseases, such as nAMD, characterized by a high dose of aflibercept and an extended dose interval.

Description

[0001] This application is related to U.S. Provisional Patent Application No. 63 / 319,869, filed March 15, 2022; U.S. Provisional Patent Application No. 63 / 404,512, filed September 7, 2022; U.S. Provisional Patent Application No. 63 / 404,893, filed September 8, 2022; U.S. Provisional Patent Application No. 63 / 411,594, filed September 29, 2022; U.S. Provisional Patent Application No. 63 / 412,165, filed September 30, 2022; and November 1, 2022. The following U.S. Provisional Patent Application No. 63 / 421,298, filed on December 22, 2022; U.S. Provisional Patent Application No. 63 / 434,920, filed on December 22, 2022; U.S. Provisional Patent Application No. 63 / 444,480, filed on February 9, 2023; and U.S. Provisional Patent Application No. 63 / 447,582, filed on February 22, 2023 and Canadian Patent Application No. 3,190,733, filed on February 22, 2023 are related to this document; each of these patent applications is incorporated herein by reference in its entirety.

[0002] References to sequence listings

[0003] This application is incorporated by reference into a computer-readable sequence list in ST.26 XML format, named 11193WO01_Sequence, created on March 13, 2023, and containing 5,464 bytes. Technical Field

[0004] The field of this invention relates to methods for treating or preventing angiogenic eye diseases by administering VEGF antagonists. Background Technology

[0005] Neovascular (wet) AMD (nAMD) is a major health problem in the global aging population. Vision loss in nAMD is caused by abnormal growth and leakage of blood vessels in the macula. Vision loss tends to have a greater impact in older subjects with nAMD because it significantly reduces visual compensation for functional impairment caused by other age-related comorbidities, such as arthritis and osteoporosis.

[0006] Intravitreal (IVT) administration of anti-vascular endothelial growth factor (VEGF) therapy (such as...) Anti-VEGF (IVT) therapy inhibits the growth and leakage of new blood vessels in the retina and is currently the standard treatment for patients with nAMD. It not only maintains visual function but also provides clinically meaningful visual gains. For most patients, treatment for nAMD is long-term and lifelong, aimed at suppressing retinal edema and recurrence of choroidal neovascularization (CNV). Although currently approved IVT therapies are effective and well-tolerated, the need for IVT injections every 4 to 8 weeks, especially during the initial and maintenance phases of treatment, places a significant burden on physicians, patients, and caregivers. While the procedure is simple and relatively easy to implement, ensuring the appropriate injection frequency to achieve patient outcomes similar to those seen in pivotal studies presents an increasing challenge for individual practice and the healthcare system as a whole. Furthermore, high-frequency dosing increases the burden on patients, such as the burden of finding transportation and disrupting work. A second effect of this burden is a reduced likelihood of non-adherence to prescribed treatment regimens.

[0007] While the efficacy and safety of currently approved VEGF antagonist therapies in treating nAMD have been proven, there remains an unmet medical need to develop therapies with the potential to reduce the treatment burden while providing visual outcomes that are at least similar to or even better than the currently available standard of care.

[0008] Increasing the molar concentration of the VEGF antagonist therapeutic protein in the dosing formulation is a potential approach to provide further benefit to subjects with choroidal retinal vascular diseases, including nAMD. Higher doses of aflibercept administered via IVT have the potential to prolong the therapeutic effect of the drug. Extending the treatment interval to every 12 or 16 weeks early after treatment initiation will reduce the number of injections during the first year of treatment. The potential reduction in injection-related treatment burden and safety events through fewer injections could be a significant contribution to patient / subject care and healthcare services. EYLEA (2 mg dose, administered at a concentration of 40 mg / mL, also known as intravitreal aflibercept [IAI]) is currently approved in the United States (US) for the treatment of nAMD and is also approved for the treatment of post-retinal vein occlusion macular edema (RVO), diabetic macular edema (DME), and diabetic retinopathy (DR). Summary of the Invention

[0009] This invention provides a method for treating or preventing neovascular age-related macular degeneration, comprising administering one or more doses (e.g., ≥8 mg) of aflibercept such that, following intravitreal injection of aflibercept, the clearance of free aflibercept from the ocular ventricle is about 0.367-0.458 mL / day (e.g., 0.41 mL / day), and the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the subject's ocular ventricle after said intravitreal injection is about 15 weeks; and the time to reach the lower limit of quantitation (LLOQ) (e.g., about 0.0156 mg / L) of free aflibercept in the subject's plasma after said intravitreal injection is about 3.5 weeks; for example, wherein the aflibercept is administered in the form of an aqueous pharmaceutical formulation, wherein the aflibercept has less than about 3.5% of high molecular weight material immediately after production and purification and / or has less than or equal to about 6% of high molecular weight material after being stored at about 2°C-8°C for about 24 months. In one embodiment of the invention, the aqueous pharmaceutical formulation comprises an aqueous pharmaceutical formulation containing: at least about 100 mg / ml of a VEGF receptor fusion protein, the fusion protein comprising two polypeptides, each polypeptide comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a polymerized component; about 10 mM-100 mM L-arginine; sucrose; a histidine-based buffer; and a surfactant; wherein the pH of the formulation is about 5.0 to about 6.8; wherein the VEGF receptor fusion protein has less than about 3.5% of high molecular weight material immediately after production and purification and / or less than or equal to about 6% of high molecular weight material after storage at about 2°C-8°C for about 24 months. In one embodiment of the invention, the method includes administering a single initial dose of about 8 mg or more of aflibercept, followed by one or more administrations of a second dose of about 8 mg or more of aflibercept, followed by one or more administrations of a third dose of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12 to 20 weeks (preferably 12, 16 or 20 weeks) immediately following the preceding dose.

[0010] This invention provides a method for achieving a clearance rate of free aflibercept from the ocular ventricle after intravitreal injection relative to intravitreal injection. <A method for slowing the clearance rate of aflibercept from the ocular ventricle after 4 mg aflibercept, comprising intravitreal injection of about 8 mg or more of a single initial dose of aflibercept into the eye of a subject in need, followed by one or more second doses of about 8 mg or more of aflibercept, followed by one or more third doses of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks after the immediate preceding dose; and wherein each third dose is administered about 12 to 20 weeks after the immediate preceding dose. In one embodiment of the invention, the clearance rate of free aflibercept from the ocular ventricle is higher than that after intravitreal injection. < The clearance rate from the ocular ventricle after 4 mg aflibercept is slow, approximately 34%. For example, after intravitreal injection of ≥8 mg aflibercept, the clearance rate of free aflibercept from the ocular ventricle is approximately 0.367–0.458 mL / day or 0.41 mL / day.

[0011] The present invention also provides a method for increasing the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the ventricle of a subject after intravitreal injection of aflibercept relative to the time to reach the LLOQ after intravitreal injection of about 2 mg of aflibercept, for example, by more than 1.3 weeks, for example, by more than 6 to 10 weeks, for example, by about 15 weeks, the method comprising intravitreal injection of a single initial dose of about 8 mg or more of aflibercept into the eye of a subject in need, followed by one or more second doses of about 8 mg or more of aflibercept, followed by one or more third doses of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 to 20 weeks after the immediately preceding dose.

[0012] The present invention also provides a method for increasing the time to reach the lower limit of quantitation (LLOQ) (e.g., about 0.0156 mg / L) of free aflibercept in the plasma of a subject after intravitreal injection of aflibercept relative to the time to reach the LLOQ of free aflibercept in the plasma of a subject after intravitreal injection of about 2 mg of aflibercept, for example, by more than 1.5 weeks, for example by about 2 weeks to about 3.5 weeks, the method comprising intravitreal injection of a single initial dose of about 8 mg or more of aflibercept into the eye of a subject in need, followed by one or more second doses of about 8 mg or more of aflibercept, followed by one or more third doses of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 to 20 weeks after the immediately preceding dose. In one embodiment of the invention, ≥8 mg of aflibercept is administered in the form of an aqueous pharmaceutical formulation comprising one or more of the following: a histidine-based buffer, arginine (e.g., L-arginine, such as L-arginine HCl), a sugar or polyol (such as sucrose), and a pH of about 5.8. In one embodiment of the invention, aflibercept immediately after production and purification has less than about 3.5% of high molecular weight material and / or less than or equal to about 6% of high molecular weight material after storage at about 2°C to 8°C for about 24 months; for example, wherein ≥8 mg of aflibercept is in an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein, the fusion protein comprising two polypeptides, each polypeptide comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a polymerized component; about 10 mM to 100 mM L-arginine; sucrose; a histidine-based buffer; and a surfactant; wherein the pH of the formulation is about 5.0 to about 6.8; wherein the VEGF receptor fusion protein immediately after production and purification has less than about 3.5% of high molecular weight material and / or less than or equal to about 6% of high molecular weight material after storage at about 2°C to 8°C for about 24 months.

[0013] This invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, for improving best-corrected visual acuity in a subject of need with nAMD, or for promoting retinal dryness in a subject of need with nAMD; the method comprising administering one or more doses of about 8 mg or more of VEGF receptor fusion protein to the eye of the subject once or multiple times every 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks, or 12-20 weeks, or 12-16 weeks, or 16-20 weeks.

[0014] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, the method comprising administering to the eye of the subject a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12 to 20 weeks immediately following the preceding dose.

[0015] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, the method comprising administering to the eye of the subject a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12 weeks immediately following the preceding dose.

[0016] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, the method comprising administering to the eye of the subject a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 16 weeks immediately following the preceding dose.

[0017] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, the method comprising administering to the eye of the subject a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediate preceding dose; and wherein each third dose is administered about 20 weeks after the immediate preceding dose.

[0018] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, comprising administering 8 mg of VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 μL), via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), followed by intravitreal injection of 8 mg aflibercept (0.07 mL) every 8 to 16 weeks (2 to 4 months + / - 7 days).

[0019] This invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, comprising administering 8 mg of VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 μL), via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter every 12 weeks (2-4 months + / - 7 days) via intravitreal injection of 8 mg of VEGF receptor fusion protein (0.07 mL).

[0020] This invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, comprising administering 8 mg of VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 μL), via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter every 16 weeks (2-4 months + / - 7 days) via intravitreal injection of 8 mg of VEGF receptor fusion protein (0.07 mL).

[0021] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, comprising administering 8 mg of VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 μL), via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter via intravitreal injection of 8 mg of VEGF receptor fusion protein (0.07 mL) every 20 weeks (+ / - 7 days).

[0022] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, wherein: (1) the subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method comprises administering the initial 8 mg dose of VEGF receptor fusion protein to the subject one month later, and administering a first 8 mg dose of VEGF receptor fusion protein one month thereafter; and administering a second 8 mg dose of VEGF receptor fusion protein one month thereafter; and then administering one or more doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen. (2) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method includes administering a first 8 mg second dose of VEGF receptor fusion protein to the subject 1 month later, followed by a second 8 mg dose of VEGF receptor fusion protein 1 month thereafter; then, every 12, 16, or 20 weeks thereafter, one or more 8 mg maintenance doses of VEGF receptor fusion protein are administered according to the HDq12, HDq16, or HDq20 dosing regimen; (3) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method includes administering a first 8 mg second dose of VEGF receptor fusion protein to the subject 1 month later, followed by a second 8 mg dose of VEGF receptor fusion protein 1 month thereafter; and then, every 12, 16, or 20 weeks thereafter, one or more 8 mg maintenance doses of VEGF receptor fusion protein are administered according to the HDq12, HDq16, or HDq20 dosing regimen. The subject was given a second dose of 8 mg of VEGF receptor fusion protein, followed by one or more doses of 8 mg maintenance VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (4) the subject had received an initial 2 mg dose of VEGF receptor fusion protein, and the method then included administering a first dose of 8 mg maintenance VEGF receptor fusion protein to the subject one month later, and all other doses of 8 mg maintenance VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; 5) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering the initial 8 mg dose of VEGF receptor fusion protein to the subject one month later, followed by a first 8 mg dose of VEGF receptor fusion protein one month later; and a second 8 mg dose of VEGF receptor fusion protein one month later; and then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.(6) The subject had received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method then included administering a first 8 mg dose of VEGF receptor fusion protein one month later, followed by a second 8 mg dose of VEGF receptor fusion protein one month thereafter; then, every 12, 16, or 20 weeks thereafter, a maintenance dose of 8 mg of VEGF receptor fusion protein was administered once or multiple times according to the HDq12, HDq16, or HDq20 dosing regimen; (7) The subject had received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method includes administering a second dose of VEGF receptor fusion protein of 2 mg once, followed by a second dose of 8 mg of VEGF receptor fusion protein one month later, and then administering one or more doses of 8 mg of VEGF receptor fusion protein every 12, 16, or 20 weeks thereafter, according to the HDq12, HDq16, or HDq20 dosing regimen; (8) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first dose of 2 mg of VEGF receptor fusion protein one month later, followed by a first dose of 8 mg of VEGF receptor fusion protein one month later, and then administering one dose of 8 mg of VEGF receptor fusion protein every 12 or 16 weeks. Or administer all other 8 mg maintenance doses of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing regimen for 20 weeks; (9) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later, then the method includes administering the initial 8 mg dose of VEGF receptor fusion protein to the subject one month later, and administering a first 8 mg dose of VEGF receptor fusion protein one month thereafter; and administering a second 8 mg dose of VEGF receptor fusion protein one month thereafter. The method comprises administering the VEGF receptor fusion protein dose once or multiple times every 12, 16 or 20 weeks thereafter, according to the HDq12 or HDq16 or HDq20 dosing regimen; (10) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein 1 month later and a second 2 mg second dose of VEGF receptor fusion protein 1 month later, and then the method comprises administering the subject a first 8 mg second dose of VEGF receptor fusion protein 1 month later and a second 8 mg second dose of VEGF receptor fusion protein 1 month thereafter;Then, every 12, 16, or 20 weeks thereafter, a maintenance dose of 8 mg of VEGF receptor fusion protein is administered once or multiple times according to the HDq12, HDq16, or HDq20 dosing regimen; (11) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later, then the method includes administering a second 8 mg dose of VEGF receptor fusion protein to the subject one month later. The method involves administering a maintenance dose of 8 mg VEGF receptor fusion protein once or multiple times every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (12) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later, and then the method includes administering a first 8 mg dose of VEGF receptor fusion protein to the subject two months later. The maintenance dose of VEGF receptor fusion protein, and every 12, 16, or 20 weeks, according to the HDq12, HDq16, or HDq20 dosing regimen, all other 8 mg maintenance doses of VEGF receptor fusion protein were administered; (13) the subject had received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later and a first 2 mg maintenance dose of VEGF after 8 weeks. The method comprises administering an initial 8 mg dose of VEGF receptor fusion protein to the subject up to 2 months after the last dose of VEGF receptor fusion protein, followed by a first 8 mg dose of VEGF receptor fusion protein 1 month thereafter; and a second 8 mg dose of VEGF receptor fusion protein 1 month thereafter; and then, once or multiple times every 12, 16, or 20 weeks, according to the HDq12, HDq16, or HDq20 dosing regimen, to maintain an 8 mg dose of VEGF receptor fusion protein.(14) The subject had received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, followed by a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later. The method then included administering a first 8 mg dose of VEGF receptor fusion protein to the subject up to two months after the last dose, followed by a second 8 mg dose of VEGF receptor fusion protein one month thereafter, and then every 12, 16, or 20 weeks thereafter. The method involves administering one or more 8 mg maintenance doses of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing regimen; (15) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later, and then the method includes administering a second 8 mg dose of VEGF receptor fusion protein to the subject up to two months after the last dose of VEGF receptor fusion protein, and then... (16) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later and a first 2 mg dose of VEGF receptor fusion protein eight weeks later, and then the method includes administering a first 8 mg maintenance dose to the subject up to two months after the last dose of VEGF receptor fusion protein. The VEGF receptor fusion protein, and every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen, all other 8 mg maintenance doses of the VEGF receptor fusion protein are administered; wherein (i) the HDq12 dosing regimen comprises: a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 weeks after the immediately preceding dose;(ii) The HDq16 dosing regimen comprises: a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 16 weeks after the immediately preceding dose; and (iii) The HDq20 dosing regimen comprises: a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 20 weeks after the immediately preceding dose—preferably, the VEGF receptor fusion protein is aflibercept, and 2 to 4 weeks is preferably 4 weeks.

[0023] The present invention also provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, wherein: (a) the subject has received an initial 8 mg dose of VEGF receptor fusion protein; the method then includes administering a first 8 mg second dose of VEGF receptor fusion protein to the subject one month later, followed by a second 8 mg dose of VEGF receptor fusion protein one month thereafter; then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to an HDq12, HDq16, or HDq20 dosing regimen; (b) the subject has received an initial 8 mg dose of VEGF receptor fusion protein... The method includes administering a second 8 mg dose of VEGF receptor fusion protein to the subject one month later, followed by administration of a second 8 mg dose of VEGF receptor fusion protein one month later, and then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (c) the subject has received an initial 8 mg dose of VEGF receptor fusion protein and received a second 8 mg dose of VEGF receptor fusion protein one month later, followed by a second 8 mg dose of VEGF receptor fusion protein one month later; the method then includes administering the second 8 mg dose of VEGF receptor fusion protein to the subject one month later. The subject was given a first 8 mg maintenance dose of VEGF receptor fusion protein after 12, 16, or 20 weeks, and all other 8 mg maintenance doses of VEGF receptor fusion protein were administered every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; or (d) wherein the subject had received an initial 8 mg dose of VEGF receptor fusion protein and received a first 8 mg second dose of VEGF receptor fusion protein one month later and a second 8 mg second dose of VEGF receptor fusion protein one month later, and thereafter, every 12, 16, or 20 weeks, the subject had received one or more 8 mg maintenance doses of VEGF receptor fusion protein. The method then comprises administering, once or more, an 8 mg maintenance dose of VEGF receptor fusion protein to the subject 12, 16, or 20 weeks after the last maintenance dose of VEGF receptor fusion protein and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; wherein, (i) the HDq12 dosing regimen comprises: a single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a third dose of about 8 mg or more of VEGF receptor fusion protein, followed by a second ... second dose of about(ii) The HDq16 dosing regimen comprises: a single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered approximately 2 to 4 weeks after the immediate preceding dose; and wherein each third dose is administered approximately 12 weeks after the immediate preceding dose. The administration regimen of HDq20 is as follows: approximately 16 weeks later; and (iii) the HDq20 dosing regimen comprises: a single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of approximately 8 mg or more of VEGF receptor fusion protein, followed by one or more third doses of approximately 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered approximately 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered approximately 20 weeks after the immediately preceding dose - preferably, the VEGF receptor fusion protein is aflibercept, and 2 to 4 weeks is preferably 4 weeks.

[0024] The present invention also provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject who has received a dosing regimen for the treatment or prevention of nAMD, the dosing regimen comprising: a single initial dose of about 2 mg of VEGF receptor fusion protein (preferably aflibercept), followed by one or more second doses of about 2 mg of VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of VEGF receptor fusion protein; wherein each second dose is administered approximately 4 weeks after the immediately preceding dose; and wherein each third dose is administered approximately 8 weeks after the immediately preceding dose; and wherein the subject is at any stage of the 2 mg VEGF receptor fusion protein dosing regimen, the method comprising administering an 8 mg dose of VEGF receptor fusion protein to the eye of the subject, evaluating the subject approximately 8, 10, or 12 weeks after said administration, and continuing to administer 8 mg every 12 or 16 weeks to the subject if the treating physician determines that dosing every 12 or 16 weeks is appropriate. VEGF receptor fusion protein; or evaluate the subject approximately 8, 10, or 12 weeks after administration, and if the treating physician determines that administration every 12 weeks is appropriate, administer another 8 mg dose of VEGF receptor fusion protein, re-evaluate the subject approximately 12 weeks later, and if the treating physician determines that administration every 16 weeks is appropriate, continue administering 8 mg of VEGF receptor fusion protein to the subject every 16 weeks.

[0025] In one embodiment of the invention, the subject has received a dosing regimen for the treatment or prevention of neovascular age-related macular degeneration: a single initial dose of about 2 mg of VEGF receptor fusion protein (preferably aflibercept), followed by two second doses of about 2 mg of VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of VEGF receptor fusion protein; wherein each second dose is administered about 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 8 weeks after the immediately preceding dose.

[0026] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need, comprising administering to the subject's eye a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12 or 16 weeks immediately following the preceding dose; the method further comprises, after receiving one or more of the third doses about 12 or 16 weeks after the preceding dose, extending the third dose interval from 12 weeks to 16 weeks; from 12 weeks to 20 weeks; or from 16 weeks to 20 weeks; for example, wherein the third dose interval is adjusted about 48 or 60 weeks after the start of treatment. In one embodiment of the invention, prior to the extension, the subject exhibits (a) BCVA loss <5 letters; and / or (b) CRT <300 or 320 μm. In another embodiment of the invention, the method further includes evaluating the subject's BVCA and / or CRT, and extending the third dose interval if the subject exhibits (a) BCVA loss <5 letters; and / or (b) CRT <300 or 320 μm.

[0027] The present invention provides a method for treating or preventing nAMD in a subject in need, the method comprising administering to the eye of the subject a single initial dose of about >8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12, 16, or 20 weeks immediately following the preceding dose; the method further comprising, after receiving one or more of the third doses about 12, 16, or 20 weeks immediately following the preceding dose, shortening the third dose interval from 12 weeks to 8 weeks; from 16 weeks to 12 weeks; from 16 weeks to 8 weeks; from 20 weeks to 8 weeks; from 20 weeks to 12 weeks; or from 20 weeks to 16 weeks. In one embodiment of the invention, prior to the shortening, the subject exhibits (a) a BCVA loss of >10 letters relative to baseline; and / or (b) a CRT increase of >50 μm relative to baseline. In another embodiment of the invention, the method further includes evaluating the subject's BVCA and / or CRT, and shortening the third dosing interval if the subject exhibits (a) a BCVA loss of >10 letters relative to baseline; and / or (b) a CRT increase of >50 μm relative to baseline.

[0028] In one embodiment of the invention, if (a) the BCVA (ETDRS) loss is greater than 5 letters relative to the BCVA observed approximately 12 weeks after treatment initiation; (b) the CRT increase is greater than 25 micrometers relative to the CRT observed approximately 12 weeks after treatment initiation; and / or (c) there is new foveal neovascularization or foveal hemorrhage; for example, at week 16 or week 20 after treatment initiation, the interval between the third doses is shortened from 12 or 16 weeks to 8 weeks; or if (a) the BCVA (ETDRS) loss is greater than 5 letters relative to the BCVA observed approximately 12 weeks after treatment initiation; (b) the CRT increase is greater than 25 micrometers relative to the CRT observed approximately 12 weeks after treatment initiation; and / or (c) there is new foveal neovascularization or foveal hemorrhage; for example, at week 24 after treatment initiation, the interval between the third doses is shortened from 16 weeks to 12 weeks.

[0029] The present invention provides a method for treating or preventing neovascular age-related macular degeneration in a subject of need, the method comprising administering three doses of about 8 mg of VEGF receptor fusion protein (preferably aflibercept) to the eye of the subject at 4-week intervals in the form of a formulation containing about 114.3 mg / ml of VEGF receptor fusion protein; and after said three doses, administering one or more doses of VEGF receptor fusion protein at intervals extended to 12 weeks, 16 weeks or 20 weeks.

[0030] The present invention provides a method for treating or preventing nAMD in a subject of need, the method comprising administering to the subject's eye a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by two second doses of about 8 mg or more of VEGF receptor fusion protein, wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and, after said doses, (a) determining whether the subject meets the requirement to reduce or extend (extend) one or more intervals between VEGF receptor fusion protein doses by 2 weeks, 3 weeks. (a) at least one criterion of 4 weeks or 2-4 weeks; and (b) if the determination is made, administer an additional dose of VEGF receptor fusion protein at the reduced or extended dose interval, wherein the criteria for reducing the interval include: 1. BCVA loss > 5 letters; 2. Central retinal thickness (CRT) increase > 25 micrometers; 3. New foveal hemorrhage; and / or 4. New foveal neovascularization, wherein the criteria for extending the interval include: 1. BCVA loss < 5 letters; 2. No fluid in the central region; 3. No new foveal hemorrhage; and / or 4. No foveal neovascularization. In one embodiment of the invention, the criteria for extending the interval include: 1. BCVA loss <5 letters from week 12; 2. No fluid in the central region on OCT; and 3. No new foveal hemorrhage or foveal neovascularization; and / or the criteria for reducing the interval include two of the following: 1. BCVA loss >5 letters from week 12; and 2. Increase in central retinal thickness (CRT) >25 micrometers or new foveal hemorrhage or new foveal neovascularization from week 12. In one embodiment of the invention, if the criteria are met, the interval is extended to 12 weeks, 16 weeks, or 20 weeks.

[0031] The present invention provides a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject who has been pretreated with one or more 2 mg doses of VEGF receptor fusion protein (preferably aflibercept). The method comprises administering a single initial dose of about 8 mg or more of VEGF receptor fusion protein to the subject's eye, followed by one or more administrations of a second dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 12 to 20 weeks (e.g., 12, 16, or 20 weeks) immediately following the preceding dose.

[0032] The present invention provides a method for treating or preventing angiogenic eye disease (preferably nAMD) in a subject of need, the method comprising administering to the subject's eye (1) a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept), followed by one or more (e.g., 2, 3 or 4) doses of about 8 mg or more of VEGF receptor fusion protein, followed by one or more doses of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) immediately following the preceding dose; and wherein each third dose is administered about 8 weeks immediately following the preceding dose; or (2) administering one or more doses of VEGF receptor fusion protein about every 4 weeks.

[0033] In one embodiment of the invention, a subject having any one or more of the following: ocular or periocular infection; active intraocular inflammation; and / or hypersensitivity reaction is excluded from treatment or prevention. It is not necessary to exclude subjects lacking this criterion. In one embodiment of the invention, the method herein further includes the step of evaluating the subject for: ocular or periocular infection; active intraocular inflammation; and / or hypersensitivity reaction; and if any one or more of these are found in the subject, the subject is excluded from treatment or prevention.

[0034] In one embodiment of the invention, adverse events in the subject are monitored, such as conjunctival hemorrhage, cataract, vitreous detachment, vitreous floaters, corneal epithelial defects, and / or increased intraocular pressure. If such adverse events (AEs) are identified, the identified AEs can be treated and / or such treatment can be discontinued or prevented.

[0035] In one embodiment of the invention, the method includes preparation prior to administration of the VEGF receptor fusion protein (preferably aflibercept). For example, the method includes, prior to each administration, providing or preparing: a single-dose glass vial with a protective plastic cap and stopper, containing approximately 70 microliters of an aqueous formulation containing 8 mg of VEGF receptor fusion protein; a filter needle, such as an 18-gauge x 1 1 / 2-inch, 5-micron filter needle, including a tip and bevel; an inventive needle, such as a 30-gauge x 1 / 2-inch injection needle; and a syringe, such as a 1-mL Luer lock syringe with a graduation marking for a 70-microliter capacity; packaged together; then (1) visually inspecting the aqueous formulation in the vial, and if particulate matter, turbidity, or discoloration is visible, using another vial containing the aqueous formulation of VEGF receptor fusion protein; (2) removing the protective plastic cap from the vial; and (3) cleaning the top of the vial with an alcohol swab; then using aseptic technique: (4) removing the 18-gauge x 1 1 / 2-inch filter needle from their packaging. (5) Attach the filter needle to the syringe by screwing it onto the tip of the Luer lock syringe; (6) Push the filter needle into the center of the stopper until it is fully inserted into the vial and the tip touches the bottom or bottom edge of the vial; (7) Draw all the contents of the VEGF receptor fusion protein vial into the syringe, keeping the vial upright and slightly tilted, while ensuring that the bevel of the filter needle is submerged in the liquid; (8) Continue to tilt the vial during the drawing process, keeping the bevel of the filter needle submerged in the preparation; (9) When the vial is emptied, pull the plunger fully back to completely empty the filter needle; (10) remove the filter needle from the syringe and dispose of it; (11) remove the 30 x 1 / 2 inch injection needle from its packaging and attach it to the syringe by firmly screwing the injection needle onto the tip of the Luer lock syringe; (12) hold the syringe with the needle facing upward and check the syringe for air bubbles, and if there are air bubbles, gently tap the syringe with your fingers until the air bubbles rise to the top; and (13) slowly press down the plunger so that the plunger tip aligns with the 70 μL mark on the syringe. In one embodiment of the invention, the injection of the VEGF receptor fusion protein is performed under controlled aseptic conditions, including surgical hand disinfection and the use of sterile gloves, sterile drapes and sterile eyelid speculum (or equivalent), and the administration of an anesthetic and a local broad-spectrum antimicrobial agent prior to injection.

[0036] In one embodiment of the invention, the subject is receiving a dosing regimen for the treatment or prevention of nAMD, which includes: a single initial dose of about 2 mg of VEGF receptor fusion protein (preferably aflibercept), followed by two second doses of about 2 mg of VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of VEGF receptor fusion protein; wherein each second dose is administered approximately 4 weeks after the preceding dose; and wherein each third dose is administered approximately 8 weeks after the preceding dose; wherein the subject is at any stage of the 2 mg VEGF receptor fusion protein dosing regimen (initial dose, second dose, or third dose).

[0037] In one embodiment of the invention, one or more second doses constitute two second doses; 2 to 4 weeks is about 4 weeks; 12-20 weeks is about 12 weeks; 12-20 weeks is about 16 weeks; 12-20 weeks is about 20 weeks; 12-20 weeks is about 12-16 weeks; 8-16 weeks is about 12 weeks; 8-16 weeks is about 16 weeks; 8-16 weeks is about 12-16 weeks; 2 to 4 weeks is about 4 weeks and one or more second doses constitute two second doses; 12-20 weeks is about 12 weeks and one or more second doses constitute two doses; 12-20 weeks is about 16 weeks and one or more second doses constitute two second doses; 12-20 weeks is about 20 weeks and one or more second doses constitute two doses; 12-20 weeks is about 12-16 weeks and one... Or multiple second doses are two second doses; 2 to 4 weeks is about 4 weeks and one or more second doses are two second doses and the VEGF receptor fusion protein is aflibercept; 12-20 weeks is about 12 weeks and one or more second doses are two second doses and the VEGF receptor fusion protein is aflibercept; 12-20 weeks is about 16 weeks and one or more second doses are two second doses and the VEGF receptor fusion protein is aflibercept; 12-20 weeks is about 20 weeks and one or more second doses are two second doses and the VEGF receptor fusion protein is aflibercept; and / or 12-20 weeks is about 12-16 weeks and one or more second doses are two second doses and the VEGF receptor fusion protein is aflibercept.

[0038] In one embodiment of the invention, the VEGF receptor fusion protein: (i) 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 130-231 of SEQ ID NO:2; and (3) a polymerized component comprising amino acids 232-457 of SEQ ID NO:2; (ii) comprises two polypeptides comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a polymerized component; (iii) comprises two polypeptides comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, an Ig domain 4 of VEGFR2, and a polymerized component; (iv) comprises two VEGFR1R2-FcΔC1(a) polypeptides encoded by the nucleic acid sequence of SEQ ID NO:1; or (v) selected from the group consisting of aflibercept and conbercept. In one embodiment of the invention, the VEGF receptor fusion protein comprises or consists of amino acids 27-457 of the amino acid sequence shown in SEQ ID NO:2.

[0039] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein (preferably aflibercept) is contained in an aqueous pharmaceutical formulation selected from the group consisting of A-KKKK.

[0040] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein (preferably aflibercept) is administered in the form of an aqueous pharmaceutical formulation containing about 114.3 mg / ml of VEGF receptor fusion protein.

[0041] In one embodiment of the invention, the VEGF receptor fusion protein (preferably aflibercept) is administered intravitreally from a syringe or pre-filled syringe, which is, for example, glass or plastic, and / or sterile. In another embodiment of the invention, the VEGF receptor fusion protein is injected intravitreally using a 30-gauge × 1 / 2-inch sterile injection needle.

[0042] In one embodiment of the invention, the subject has previously received one or more doses of 2 mg VEGF receptor fusion protein (preferably aflibercept), such as Eylea.

[0043] In one embodiment of the invention, the method described herein further includes administering one or more additional doses.

[0044] In one embodiment of the invention, 2 mg of VEGF receptor fusion protein (preferably aflibercept) is in an aqueous pharmaceutical formulation containing 40 mg / ml of VEGF receptor fusion protein, for example containing 40 mg / ml of VEGF receptor fusion protein, 10 mM sodium phosphate, 40 mM NaCl, 0.03% polysorbate 20 and 5% sucrose, at a pH of 6.2.

[0045] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising ≥100 mg / ml VEGF receptor fusion protein (preferably aflibercept), a histidine-based buffer, and arginine (e.g., L-arginine). In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising a sugar or polyol, such as sucrose. In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation having a pH of about 5.8. In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising about 103-126 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine, for example, comprising about 114.3 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0046] In embodiments of the present invention, 8 mg of the VEGF antagonist is administered in the following doses: 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; 71μl; 72μl; 73μl; 74μl; 75μl; 76μl; 77μl; 78μl; 79μl; 80μl; 81μl; 82μl; 83μl; 84μl; 85μl; 86μl; 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, about 70 ± 4 or 5 μl.

[0047] In one embodiment of the invention, the method herein includes the step of administering a VEGF receptor fusion protein (preferably aflibercept) to the eyes of a subject, for example, via intravitreal administration.

[0048] In one embodiment of the invention, the subject achieves and / or maintains one or more of the following: an improvement in best-corrected visual acuity (BCVA) of ≥5, ≥10, ≥15, or ≥20 letters; no decrease in best-corrected visual acuity (BCVA); elimination of retinal fluid; elimination of intraretinal fluid (IRF) and / or subretinal fluid; reduction in total damaged choroidal neovascularization (CNV) area; loss or reduction of intraretinal fluid; loss or reduction of subretinal fluid; reduction in central retinal thickness (CST); improvement in vision-related quality of life; absence of treatment-related adverse events (AEs) and / or serious adverse events (SAEs); and an ETDRS letter score of at least 69 (approximately 20 / 40 Snellen equivalents). Early Treatment Diabetic Retinopathy Study (ETDRS) criteria include: an increase in BCVA (Browser Capacity Deficiency) measured by the visual acuity chart of ≥5, ≥10, ≥15, or ≥20 letters, or no loss during the course of treatment; an increase in mean BCVA over a 12-week period; absence of intraretinal fluid (IRF) and subretinal fluid; reduced choroidal neovascularization (CNV) size; reduced total damaged CNV area compared to baseline; loss of IRF and / or SRF; reduced central retinal thickness (CST); improved vision-related quality of life as measured by the National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25); absence of adverse events (AEs) and / or serious AEs (SAEs) occurring during the treatment period; and [further criteria for criteria in subjects with nAMD]. The efficacy and / or safety in this study were similar to those of aflibercept administered intravitreally at approximately 2 mg every 4 weeks for the first 3 months, followed by approximately 2 mg intravitreally every 8 weeks or every 2 months, where efficacy was measured as an increase in BCVA and / or a decrease in central retinal thickness, and where safety was measured as the incidence of adverse events such as increased blood pressure, increased intraocular pressure, visual disturbances, vitreous floaters, vitreous detachment, iris neovascularization, and / or vitreous hemorrhage; no anti-drug antibodies were detected during treatment; from the start of treatment (baseline) to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and 48 Improvement in best-corrected visual acuity (BVCA) at week 1, week 52, week 56, or week 60; an increase in BCVA of ≥2, ≥3, ≥4, ≥5, ≥6, or ≥7 letters as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent; an improvement in BCVA of approximately 2 or 3 letters (ETDRS or Snellen equivalent) at 4 weeks after treatment initiation with the HDq12 regimen; or approximately 3 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; an improvement in BCVA of approximately 5 letters (ETDRS or Snellen equivalent) at 8 weeks after treatment initiation with the HDq12 regimen.Or approximately 4 or 5 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 12 weeks after treatment initiation is approximately 5 or 6 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 16 weeks after treatment initiation is approximately 6 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 20 weeks after treatment initiation is approximately [missing information - likely a number]. Approximately 6 letters (ETDRS or Snellen equivalent); or approximately 6 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 24 weeks after treatment initiation is approximately 5 or 6 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 28 weeks after treatment initiation is approximately 7 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; at 32 weeks after treatment initiation The improvement in BCVA at 36 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; and approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen. The improvement in BCVA at 40 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen. The improvement in BCVA at 44 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; the improvement in BCVA at 48 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; and the improvement in BCVA at 52 weeks after treatment initiation was approximately 7 or 8 letters (ETDRS or Snellen equivalent) with the HDq12 regimen.Or approximately 6 or 7 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 56 weeks after treatment initiation with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 60 weeks after treatment initiation with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at approximately 48 to 60 weeks after treatment initiation with the HDq12 regimen. Approximately 6 or 7 letters (ETDRS or Snellen equivalent); or approximately 6 or 7 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; BCVA improvement at 60 weeks after treatment initiation is approximately 5, 10, or 15 letters (ETDRS or Snellen equivalent) when using the HDq12 or HDq16 regimen; BCVA improvement at approximately weeks 8, 9, 10, 11, or 12 after treatment initiation, and thereafter maintained during the treatment regimen (within approximately ±1 or ±2 ETDRS letters or Snellen equivalents); BCVA at 4 weeks after treatment initiation is approximately 63 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or When using the HDq16 regimen, the BCVA is approximately 63 letters (ETDRS or Snellen equivalent); at 8 weeks post-treatment initiation, the BCVA is approximately 65 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or at 12 weeks post-treatment initiation, the BCVA is approximately 66 letters (ETDRS or Snellen equivalent); or at 16 weeks post-treatment initiation, the BCVA is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen. Approximately 66 letters (ETDRS or Snellen equivalent); or approximately 66 letters (ETDRS or Snellen equivalent) in the HDq16 regimen; approximately 66 letters (ETDRS or Snellen equivalent) in the HDq12 regimen at 20 weeks after treatment initiation; or approximately 66 letters (ETDRS or Snellen equivalent) in the HDq16 regimen; approximately 66 letters (ETDRS or Snellen equivalent) in the HDq12 regimen at 24 weeks after treatment initiation; or approximately 66 letters (ETDRS or Snellen equivalent) in the HDq16 regimen.The BCVA at 28 weeks after treatment initiation was approximately 67 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; the BCVA at 32 weeks after treatment initiation was approximately 67 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; the BCVA at 36 weeks after treatment initiation was approximately 67 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) with the HDq16 regimen. The BCVA is approximately 66 letters (ETDRS or Snellen equivalent) at 40 weeks post-treatment initiation with the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; approximately 68 letters (ETDRS or Snellen equivalent) at 44 weeks post-treatment initiation with the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; and approximately 67 letters (ETDRS or Snellen equivalent) at 48 weeks post-treatment initiation with the HDq12 regimen. The BCVA at 52 weeks post-treatment is approximately 67 or 68 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen. The BCVA at 60 weeks after treatment initiation was approximately 66 or 67 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; the BCVA between approximately 48 and approximately 60 weeks after treatment initiation was approximately 66 to approximately 72 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 66 to approximately 70 letters (ETDRS or Snellen equivalent) with the HDq16 regimen; and the retina was free of fluid in the central region (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]).No subretinal pigment epithelial fluid; no fluid leakage on fluorescein angiography (FA); a reduction in central retinal thickness (CRT) of at least approximately 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 micrometers; a change in central retinal thickness 4 weeks after treatment initiation of approximately -120, -121, -122, -122.4, or -120.2 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 regimen; or approximately -126, -127, -126.6, or -126.3 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 regimen; treatment The change in central retinal thickness at 8 weeks after treatment initiation was approximately -132, -133, -134, -135, -136, -136.2, or -132.8 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -139, -140, -139.5, or -139.6 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen. The change in central retinal thickness at 12 weeks after treatment initiation was approximately -136, -137, -138, -139, -140, -141, -140.9, or -136.6 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -139, -140, -139, -140, -141, -140.9, or -136.6 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen. The initial retinal thickness was approximately -144, -143, or -143.5 micrometers (±10, 11, or 12 micrometers); the change in central retinal thickness at 16 weeks after treatment initiation was approximately -120, -121, -122, -123, -124, -123.4, or -120.1 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -132, -133, -132.1, or -133.1 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen; the change in central retinal thickness at 20 weeks after treatment initiation was approximately -110, -111, -112, -113, -114, or -113 with the HDq12 regimen. 6 or -110.9 micrometers (±10, 11 or 12 micrometers); or approximately -115 or -116 or -117 or -118 or -115.8 or -117.7 micrometers (±10, 11 or 12 micrometers) when using the HDq16 regimen; changes in central retinal thickness 24 weeks after treatment initiation are approximately -134, -135, -136 or -137 or -138 or -137.6 or -134.9 micrometers (±10, 11 or 12 micrometers) when using the HDq12 regimen; or approximately -105 or -106 or -107 or -108 or -105.3 or -107.8 micrometers (±10, 11 or 12 micrometers) when using the HDq16 regimen;The change in central retinal thickness at 28 weeks after treatment initiation was approximately -130, -131, -132, -133, -134, -133.7, or -130.7 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -144, -145, -146, -147, -148, -144.7, or -147.2 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen; the change in central retinal thickness at 32 weeks after treatment initiation was approximately -118, -19, -120, -121, -120.4, or -118.1 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -144, -145, -146, -147, -148, -144.7, or -147.2 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen; or approximately -118.1 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -118.1 micrometers (±10, 11, or 12 micrometers) with the HDq16 ...2 regimen; or approximately -118.1 micrometers (±10, 11, or 12 micro The central retinal thickness change at 36 weeks after treatment initiation was approximately -141, -142, -143, -144, -141.5, or -144 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen; at the HDq12 regimen, the central retinal thickness change was approximately -142, -143, -144, -144.2, or -142.2 micrometers (±10, 11, or 12 micrometers); or at the HDq16 regimen, the central retinal thickness change was approximately -126, -127, -128, -129, -130, -131, -126.4, or -130.5 micrometers (±10, 11, or 12 micrometers); at the HDq12 regimen, the central retinal thickness change was approximately... -131, -132, -133, or -134, or -133.8, or -131.2 micrometers (±10, 11, or 12 micrometers); or approximately -127, -128, or -127.5 micrometers (±10, 11, or 12 micrometers) when using the HDq16 regimen; changes in central retinal thickness at 44 weeks after treatment initiation are approximately -120, -121, -122, -123, -124, -125, -124.7, or -120.3 micrometers (±10, 11, or 12 micrometers) when using the HDq12 regimen; or approximately -143, -144, -145, or -144.8 micrometers (±10, 11, or 12 micrometers) when using the HDq16 regimen; The change in central retinal thickness at 48 weeks after treatment initiation was approximately -142, -143, -144, -144.4, or -142.3 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -143, -144, -145, -146, -147, -148, -143.8, or -147.1 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen; the change in central retinal thickness at 52 weeks after treatment initiation was approximately -143.2 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -139.6 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen.The change in central retinal thickness at 56 weeks after treatment initiation was approximately -136.3 μm (±10, 11, or 12 μm) with the HDq12 regimen and approximately -137.5 μm (±10, 11, or 12 μm) with the HDq16 regimen. The change in central retinal thickness at 60 weeks after treatment initiation was approximately -151.8 μm (±10, 11, or 12 μm) with the HDq12 regimen and approximately -148.8 μm (±10, 11, or 12 μm) with the HDq16 regimen. The central retinal thickness at week 4 after treatment initiation was approximately 248.2 μm with the HDq12 regimen and approximately 244.1 μm with the HDq16 regimen. At week 8 post-treatment, the central retinal thickness was approximately 234.4 μm with the HDq12 regimen and approximately 231.2 μm with the HDq16 regimen; at week 12 post-treatment, the central retinal thickness was approximately 229.7 μm with the HDq12 regimen and approximately 226.7 μm with the HDq16 regimen; at week 16 post-treatment, the central retinal thickness was approximately 247.2 μm with the HDq12 regimen and approximately 238.6 μm with the HDq16 regimen; at week 20 post-treatment, the central retinal thickness was approximately 257 μm with the HDq12 regimen and approximately 254.9 μm with the HDq16 regimen; treatment At week 24 after treatment initiation, the central retinal thickness was approximately 233 μm with the HDq12 regimen and approximately 265.4 μm with the HDq16 regimen; at week 28 after treatment initiation, the central retinal thickness was approximately 236.9 μm with the HDq12 regimen and approximately 226 μm with the HDq16 regimen; at week 32 after treatment initiation, the central retinal thickness was approximately 250.2 μm with the HDq12 regimen and approximately 229.2 μm with the HDq16 regimen; at week 36 after treatment initiation, the central retinal thickness was approximately 226.4 μm with the HDq12 regimen and approximately 244.3 μm with the HDq16 regimen; The central retinal thickness at week 40 after treatment initiation was approximately 236.8 μm with the HDq12 regimen and approximately 243.7 μm with the HDq16 regimen; the central retinal thickness at week 44 after treatment initiation was approximately 245.9 μm with the HDq12 regimen and approximately 227.7 μm with the HDq16 regimen; the central retinal thickness at week 48 after treatment initiation was approximately 226.2 μm with the HDq12 regimen and approximately 226.9 μm with the HDq16 regimen; the central retinal thickness at week 52 after treatment initiation was approximately 227.4 μm with the HDq12 regimen and approximately 231.1 μm with the HDq16 regimen.At week 56 after treatment initiation, the central retinal thickness was approximately 234.3 μm with the HDq12 regimen and approximately 233.2 μm with the HDq16 regimen; at week 60 after treatment initiation, the central retinal thickness was approximately 218.8 μm with the HDq12 regimen and approximately 221.9 μm with the HDq16 regimen; the CRT decreased at approximately weeks 4, 5, 6, 7, or 8 after treatment initiation and remained stable thereafter during the treatment regimen (within approximately ±10, ±11, or ±12 μm); approximately 4 hours after the initiation of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.0409 (± 0.0605 mg / L or 0 mg / L (or <0.0156 mg / L); approximately 8 hours after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.05 (±3.78), 0.0973 (±0.102), or 0.0672 mg / L; approximately day 2 after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.11 (±2.21), 0.146 (±0.110), or 0.0903 mg / L; approximately day 3 after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.11 (±2.06), 0.137 (±0.0947), or 0.112 mg / L. g / L; around day 5 after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.08 (±1.86), 0.0933 (±0.0481), or 0.0854 mg / L; around day 8 after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.07 (±1.75), 0.0794 (±0.0413), or 0.0682 mg / L; around day 15 after the start of HDq12 or HDq16 treatment, the plasma free aflibercept concentration was approximately 0.04 (±1.76), 0.0435 (±0.0199), or 0.0385 mg / L; Approximately day 22 after treatment initiation, the plasma concentration of free aflibercept was approximately 0.02 (±1.76), 0.0213 (±0.0148), or 0.0232 mg / L; approximately day 29 after treatment initiation with HDq12 or HDq16, the plasma concentration of free aflibercept was approximately 0.00766 (±0.00958) or 0 mg / L (or <0.0156 mg / L); approximately 4 hours after administration of HDq12 or HDq16 treatment, the plasma concentration of adjusted bound aflibercept was approximately 0.00 mg / L; approximately 8 hours after administration of HDq12 or HDq16 treatment, the plasma concentration of adjusted bound aflibercept was approximately 0.00 mg / L.On approximately day 2 following the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.06 (±3.50) or 0.124 (±0.186) mg / L; on approximately day 3 following the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.13 (±2.07) or 0.173 (±0.155) mg / L; on approximately day 5 following the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.18 (±1.88) or 0.223 (±0.157) mg / L; after the initiation of HDq12 or HDq16 treatment... Around day 8, the adjusted plasma concentration of bound aflibercept was approximately 0.31 (±1.56) or 0.334 (±0.135) mg / L; around day 15 after initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.37 (±1.50) or 0.393 (±0.130) mg / L; around day 22 after initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.25 (±3.00) or 0.335 (±0.155) mg / L; around day 29 after initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately... The combined aflibercept concentration was approximately 0.32 (±1.39) or 0.331 (±0.0953) mg / L; non-inferiority in BVCA compared to aflibercept administered intravitreally at approximately 2 mg every 4 weeks for the first 3, 4, or 5 injections, followed by approximately 2 mg every 8 weeks or every 2 months; ocular and non-ocular safety or mortality in subjects with DME was similar to aflibercept administered intravitreally at approximately 2 mg every 4 weeks for 5 injections, followed by approximately 2 mg every 8 weeks or every 2 months; and the best corrected visual acuity improved from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48. Good; from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60, the best corrected visual acuity increases by ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or ≥7 letters as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent; from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60, the retina has no fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the central zone;And / or from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60, the central retinal thickness (CRT) is reduced by at least 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 micrometers. In one embodiment of the invention, dry retina lacks intraretinal fluid and / or subretinal fluid; or dry retina is characterized by the absence of intraretinal fluid (IRF) and subretinal fluid (SRF) in the subject's eye. In one embodiment of the invention, dry retina is characterized by the absence of intraretinal fluid (IRF) and subretinal fluid (SRF) in the subject's eye after the subject has received three months' worth of VEGF receptor fusion protein.

[0049] In one embodiment of the invention, the subject is given one initial dose, two second doses, and three third doses in the first year; or the subject is given one initial dose, two second doses, and two third doses in the first year; or the subject is given one initial dose, two second doses, and three third doses in the first year, followed by two to four third doses in the second year.

[0050] In one embodiment of the invention, based on visual and / or anatomical results, such as according to Figure 3 and / or Figure 4 Adjust (increase / maintain / decrease) the dose interval according to the standard shown.

[0051] In one embodiment of the invention, 12-20 weeks refers to 12, 13, 14, 15, 16, 17, 18, 19 or 20 weeks; while 2-4 weeks refers to 2, 3, 4 or 5 weeks.

[0052] Preferably, the VEGF receptor fusion protein in this article is aflibercept.

[0053] This invention provides a kit comprising: a container containing VEGF receptor fusion protein (preferably aflibercept); and instructions for use of the VEGF receptor fusion protein, wherein the container is a vial or a pre-filled syringe, wherein the container contains ≥100 mg / mL of VEGF receptor fusion protein, wherein the container contains ≥114.3 mg / mL of VEGF receptor fusion protein, wherein the instructions include instructions for administering the VEGF receptor fusion protein to patients with DME / AMD, wherein the instructions include instructions for starting treatment with 8 mg of VEGF receptor fusion protein once a month (once every 4 weeks) for 3 consecutive doses, wherein the instructions include instructions for extending the injection interval to every 16 weeks or every 20 weeks after the initial 3 consecutive doses, and wherein the instructions include instructions for adjusting the treatment interval based on the physician's judgment of visual and / or anatomical results.

[0054] This invention provides aflibercept for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, the treatment or prevention comprising administering one or more doses of aflibercept at intervals and in quantities such that, following intravitreal injection of aflibercept, the clearance of free aflibercept from the ocular ventricle is approximately 0.37-0.46 mL / day, the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the subject's ocular ventricle following the intravitreal injection is approximately 15 weeks, and the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the subject's plasma following the intravitreal injection is approximately 3.5 weeks.

[0055] This invention provides aflibercept for use in a method for reducing the clearance rate of free aflibercept from the ocular ventricle after intravitreal injection relative to the rate of clearance of aflibercept from the ocular ventricle after intravitreal injection of <4 mg aflibercept, wherein the method comprises intravitreal injection of a single initial dose of about 8 mg or more of aflibercept into the eye of a subject in need, followed by one or more second doses of about 8 mg or more of aflibercept, followed by one or more third doses of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 to 20 weeks after the immediately preceding dose.

[0056] This invention provides aflibercept used in a method for increasing the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the eye chamber of a subject after intravitreal injection of aflibercept relative to the time to reach the LLOQ after intravitreal injection of about 2 mg of aflibercept, wherein the method comprises intravitreal injection of about 8 mg or more of a single initial dose of aflibercept into the eye of a subject in need, followed by one or more second doses of about 8 mg or more of aflibercept, followed by one or more third doses of about 8 mg or more of aflibercept; wherein each second dose is administered about 2 to 4 weeks immediately following the preceding dose; and

[0057] Each third dose is administered approximately 12-20 weeks after the preceding dose.

[0058] This invention provides a method for increasing the time to reach the lower limit of quantification (LLOQ) of free aflibercept in a subject's plasma after intravitreal injection of aflibercept, relative to the time to reach the LLOQ of free aflibercept in a subject's plasma after intravitreal injection of approximately 2 mg of aflibercept.

[0059] This method involves injecting the vitreous humor into the eye of the recipient.

[0060] A single initial dose of approximately 8 mg or more of aflibercept, followed by

[0061] A second dose of aflibercept, once or multiple times, approximately 8 mg or more, followed by

[0062] A third dose of aflibercept, once or multiple times, approximately 8 mg or more;

[0063] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and each third dose is administered approximately 12 to 20 weeks after the preceding dose.

[0064] This invention provides a VEGF receptor fusion protein used in a method described below for the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need.

[0065] - Used to improve the best corrected visual acuity of subjects in need with nAMD; or

[0066] - To promote retinal dryness in needy subjects with nAMD;

[0067] The method involves administering one or more doses of approximately 8 mg or more of the VEGF receptor fusion protein to the eyes of the subject every 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks, or 12-20 weeks, or 12-16 weeks, or 16-20 weeks.

[0068] This invention provides aflibercept for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein the treatment or prevention comprises initiating treatment with an 8 mg aflibercept injection once monthly (every 4 weeks) for three consecutive doses, followed by one or more injections every 8-16 weeks or 8-20 weeks, wherein the concentration of aflibercept at each dose is 114.3 mg / mL or wherein the application volume of each dose is 70 μL. In one embodiment of the invention, the treatment interval between two subsequent administrations of ≥8 mg aflibercept is adjusted (increased / maintained / decreased) based on visual and / or anatomical results, such as, but not limited to, an increase or loss of letters in BCVA; an increase or decrease in CRT; the presence of subretinal fluid; or the presence of hemorrhage. In one embodiment of the invention, if the subject has been identified as meeting at least one of the following criteria for reducing the treatment interval, the treatment interval is reduced by 2-4 weeks, 2 weeks, 3 weeks, or 4 weeks compared to the previous treatment interval: BCVA loss > 5 letters; central retinal thickness (CRT) increase > 25 micrometers; new foveal hemorrhage; or new foveal neovascularization. In one embodiment of the invention, if the subject has been identified as meeting at least one of the following criteria for extending the treatment interval, the treatment interval is extended by 2-4 weeks, 2 weeks, 3 weeks, or 4 weeks compared to the previous treatment interval: BCVA loss < 5 letters; no fluid in the central region; no new foveal hemorrhage; or no foveal neovascularization.

[0069] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein the method comprises administering 8 mg of VEGF receptor fusion protein (0.07 mL or 70 μL) via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), followed by intravitreal injection of 8 mg of VEGF receptor fusion protein (0.07 mL) every 8-16 weeks (2-4 months + / - 7 days) or every 8-20 weeks (2-5 months + / - 7 days).

[0070] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, wherein the method comprises administering 8 mg of VEGF receptor fusion protein (0.07 mL or 70 μL) via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter every 12 weeks (2-4 months + / - 7 days) via intravitreal injection.

[0071] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein the method comprises administering 8 mg of VEGF receptor fusion protein (0.07 mL or 70 μL) via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter every 16 weeks (2-4 months + / - 7 days) via intravitreal injection.

[0072] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of disease in subjects in need, wherein the method comprises administering 8 mg of VEGF neovascular age-related macular degeneration (nAMD) receptor fusion protein (0.07 mL or 70 μL) via intravitreal injection for the first three doses every 4 weeks (approximately every 28 days + / - 7 days, once a month), and thereafter every 20 weeks (2-4 months + / - 7 days) via intravitreal injection of 8 mg of VEGF receptor fusion protein (0.07 mL).

[0073] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein:

[0074] (1) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method includes administering an initial ≥8 mg dose of VEGF receptor fusion protein to the subject after 1 month, followed by a first ≥8 mg dose of VEGF receptor fusion protein after 1 month thereafter; and a second ≥8 mg dose of VEGF receptor fusion protein after 1 month thereafter; and then, once or more every 12, 16 or 20 weeks thereafter, according to the HDq12 or HDq16 or HDq20 dosing regimen, a maintenance dose of ≥8 mg of VEGF receptor fusion protein is administered.

[0075] (2) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method includes administering a first ≥8 mg second dose of VEGF receptor fusion protein to the subject after 1 month, and administering a second ≥8 mg second dose of VEGF receptor fusion protein after 1 month thereafter; and then, every 12, 16 or 20 weeks thereafter, administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0076] (3) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and then the method includes administering a second dose of ≥8 mg of VEGF receptor fusion protein to the subject after 1 month, and then, thereafter, administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen;

[0077] (4) The subject has received an initial 2 mg dose of VEGF receptor fusion protein, and then the method includes administering the subject a first ≥8 mg maintenance dose of VEGF receptor fusion protein after 1 month, and administering all other ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0078] (5) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering the initial ≥8 mg dose of VEGF receptor fusion protein to the subject one month later, followed by a first ≥8 mg dose of VEGF receptor fusion protein one month later; and administering a second ≥8 mg dose of VEGF receptor fusion protein one month later; and then administering one ≥8 mg dose of VEGF receptor fusion protein one or more times every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.

[0079] (6) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering a first ≥8 mg dose of VEGF receptor fusion protein to the subject one month later, followed by a second ≥8 mg dose of VEGF receptor fusion protein one month thereafter; and then, every 12, 16, or 20 weeks thereafter, administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing regimen.

[0080] (7) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later. The method then includes administering a second dose of ≥8 mg of VEGF receptor fusion protein to the subject one month later, and then administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16 or 20 weeks thereafter, according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0081] (8) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering the subject a first ≥8 mg maintenance dose of VEGF receptor fusion protein one month later, and administering all other ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.

[0082] (9) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering the initial ≥8 mg dose of VEGF receptor fusion protein to the subject one month later, and administering a first ≥8 mg dose of VEGF receptor fusion protein one month thereafter; and administering a second ≥8 mg dose of VEGF receptor fusion protein one month thereafter; and then administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.

[0083] (10) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering a first ≥8 mg dose of VEGF receptor fusion protein to the subject one month later, and administering a second ≥8 mg dose of VEGF receptor fusion protein one month thereafter; then, thereafter, every 12, 16, or 20 weeks, administer one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing regimen.

[0084] (11) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later and a second 2 mg second dose of VEGF receptor fusion protein one month later. The method then includes administering a second ≥8 mg second dose of VEGF receptor fusion protein to the subject one month later, and then administering one or more ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16 or 20 weeks thereafter, according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0085] (12) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later and a second 2 mg dose of VEGF receptor fusion protein one month later. The method then includes administering the subject a first ≥8 mg maintenance dose of VEGF receptor fusion protein two months later, and administering all other ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.

[0086] (13) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later. The method then includes administering the initial ≥8 mg dose of VEGF receptor fusion protein to the subject up to two months after the last dose of VEGF receptor fusion protein, and administering a first ≥8 mg dose of VEGF receptor fusion protein one month thereafter; and administering a second ≥8 mg dose of VEGF receptor fusion protein one month thereafter; and then administering one or more ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen.

[0087] (14) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later. The method then includes administering the subject a first ≥8 mg dose of VEGF receptor fusion protein up to two months after the last dose of VEGF receptor fusion protein, and a second ≥8 mg dose of VEGF receptor fusion protein one month thereafter, and then administering one or more ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0088] (15) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later, and then the method includes administering a second dose of ≥8 mg of VEGF receptor fusion protein to the subject up to two months after the last dose of VEGF receptor fusion protein, and then thereafter administering one or more doses of ≥8 mg maintenance dose of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen;

[0089] (16) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and a first 2 mg dose of VEGF receptor fusion protein one month later, and a second 2 mg dose of VEGF receptor fusion protein one month later, and a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later, and then the method includes administering the subject a first ≥8 mg maintenance dose of VEGF receptor fusion protein up to two months after the last dose of VEGF receptor fusion protein, and administering all other ≥8 mg maintenance doses of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen;

[0090] in,

[0091] (i) The HDq12 dosing regimen includes:

[0092] A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by...

[0093] One or more second doses of the VEGF receptor fusion protein, approximately 8 mg or more, followed by

[0094] One or more doses of approximately 8 mg or more of the VEGF receptor fusion protein;

[0095] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0096] Each third dose is administered approximately 12 weeks after the preceding dose.

[0097] (ii) The HDq16 dosing regimen includes:

[0098] A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by...

[0099] One or more second doses of the VEGF receptor fusion protein, approximately 8 mg or more, followed by

[0100] One or more doses of approximately 8 mg or more of the VEGF receptor fusion protein;

[0101] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0102] Each third dose is administered approximately 16 weeks after the preceding dose.

[0103] and

[0104] (iii) The HDq20 dosing regimen includes:

[0105] A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by...

[0106] One or more second doses of the VEGF receptor fusion protein, approximately 8 mg or more, followed by

[0107] One or more doses of approximately 8 mg or more of the VEGF receptor fusion protein;

[0108] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0109] Each third dose is administered approximately 20 weeks after the preceding dose.

[0110] This invention provides aflibercept for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, the treatment or prevention comprising administering a single initial dose of aflibercept ≥8 mg to the subject's eye, followed by one or more third doses of aflibercept of about ≥8 mg; wherein each third dose is administered about 8, 12, 16, or 20 weeks immediately following the preceding dose. In one embodiment of the invention, the subject is not a treatment-naïve subject, or the subject has been pretreated with a VEGF antagonist, or preferably the subject has been pretreated with 8 mg or 2 mg of aflibercept.

[0111] This invention provides aflibercept for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in a subject who is pre-treated with 2 mg of aflibercept. This treatment or prevention comprises administering a single initial dose of about ≥8 mg of aflibercept to the subject's eye, followed by one or more administrations of a second dose of about ≥8 mg of aflibercept, followed by one or more administrations of a third dose of about 8 mg of aflibercept, wherein each second dose is administered approximately 4 weeks after the immediately preceding dose and each third dose is administered approximately 8, 10, 12, 14, 16, 18, or 20 weeks after the immediately preceding dose. In one embodiment of the invention, one or more doses of 8 mg aflibercept are administered to the subject's eye according to HDq12, HDq16, HDq20, or a treatment and extended dosing regimen.

[0112] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects who have received a dosing regimen for the treatment or prevention of said condition, wherein:

[0113] (a) The subject has received an initial dose of ≥8 mg of VEGF receptor fusion protein, and the method comprises administering a first dose of ≥8 mg of VEGF receptor fusion protein to the subject after 1 month and administering a second dose of ≥8 mg of VEGF receptor fusion protein after 1 month thereafter; and then, thereafter, administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0114] or

[0115] (b) The subject has received an initial dose of ≥8 mg of VEGF receptor fusion protein and received a first dose of ≥8 mg of VEGF receptor fusion protein one month later. The method then includes administering a second dose of ≥8 mg of VEGF receptor fusion protein to the subject one month later, and then administering one or more doses of ≥8 mg of VEGF receptor fusion protein every 12, 16 or 20 weeks thereafter, according to the HDq12 or HDq16 or HDq20 dosing regimen.

[0116] or

[0117] (c) The subject has received an initial dose of ≥8 mg of VEGF receptor fusion protein and received a first dose of ≥8 mg of VEGF receptor fusion protein one month later and a second dose of ≥8 mg of VEGF receptor fusion protein one month later, and then the method includes administering the subject a first dose of ≥8 mg of VEGF receptor fusion protein after 12, 16 or 20 weeks, and administering all other doses of ≥8 mg of VEGF receptor fusion protein every 12, 16 or 20 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen;

[0118] or

[0119] (d) wherein the subject has received an initial dose of ≥8 mg of VEGF receptor fusion protein and a first dose of ≥8 mg of VEGF receptor fusion protein one month later and a second dose of ≥8 mg of VEGF receptor fusion protein one month later, and thereafter the subject has received one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16, or 20 weeks; and then the method comprises administering one or more maintenance doses of ≥8 mg of VEGF receptor fusion protein to the subject 12, 16, or 20 weeks after the last maintenance dose of VEGF receptor fusion protein and administering all other maintenance doses of ≥8 mg of VEGF receptor fusion protein every 12, 16, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen;

[0120] in,

[0121] (i) The HDq12 dosing regimen includes:

[0122] A single initial dose of approximately ≥8 mg or more of VEGF receptor fusion protein, followed by

[0123] One or more second doses of the VEGF receptor fusion protein of approximately ≥8 mg or more, followed by

[0124] A third dose of the VEGF receptor fusion protein, administered once or multiple times, approximately ≥8 mg or more;

[0125] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0126] Each third dose is administered approximately 12 weeks after the preceding dose.

[0127] (ii) The HDq16 dosing regimen includes:

[0128] A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by...

[0129] One or more second doses of the VEGF receptor fusion protein, approximately 8 mg or more, followed by

[0130] One or more doses of approximately 8 mg or more of the VEGF receptor fusion protein;

[0131] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0132] Each third dose is administered approximately 16 weeks after the preceding dose.

[0133] and

[0134] (iii) The HDq20 dosing regimen includes:

[0135] A single initial dose of approximately 8 mg or more of VEGF receptor fusion protein, followed by...

[0136] One or more second doses of the VEGF receptor fusion protein, approximately 8 mg or more, followed by

[0137] One or more doses of approximately 8 mg or more of the VEGF receptor fusion protein;

[0138] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and

[0139] Each third dose is administered approximately 20 weeks after the preceding dose.

[0140] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in a subject who has received a dosing regimen for the treatment or prevention of nAMD, the dosing regimen comprising: a single initial dose of about 2 mg of VEGF receptor fusion protein, followed by one or more second doses of about 2 mg of VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of VEGF receptor fusion protein; wherein each second dose is administered approximately 4 weeks after the immediately preceding dose; and wherein each third dose is administered approximately 8 weeks after the immediately preceding dose; and wherein the subject is at any stage of the 2 mg VEGF receptor fusion protein dosing regimen, the method comprising administering a dose of ≥8 mg of VEGF receptor fusion protein to the eye of the subject, evaluating the subject approximately 8, 10, or 12 weeks after said administration, and continuing to administer ≥8 mg every 12 or 16 weeks if the treating physician determines that dosing every 12 or 16 weeks is appropriate. VEGF receptor fusion protein; or evaluate the subject approximately 8, 10, or 12 weeks after administration, and if the treating physician determines that administration every 12 weeks is appropriate, administer another dose of ≥8 mg of VEGF receptor fusion protein, re-evaluate the subject within approximately 12 weeks, and if the treating physician determines that administration every 16 weeks is appropriate, continue administering 8 mg of VEGF receptor fusion protein to the subject every 16 weeks.

[0141] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, wherein the treatment or prevention comprises administering a single initial dose of about 8 mg or more of the VEGF receptor fusion protein to the subject's eye, followed by one or more administrations of a second dose (preferably two doses) of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks immediately following the preceding dose; and wherein each third dose is administered about 12 or 16 weeks immediately following the preceding dose;

[0142] The method further includes extending the third dose interval after receiving one or more third doses approximately 12 or 16 weeks after the preceding dose.

[0143] • Extended from 12 weeks to 16 weeks;

[0144] • Extended from 12 weeks to 20 weeks; or

[0145] • Extended from 16 weeks to 20 weeks

[0146] This is immediately after the previous dose.

[0147] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, the treatment and prevention comprising administering a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more administrations of a second dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks immediately following the preceding dose; and wherein each third dose is administered about 12, 16, or 20 weeks immediately following the preceding dose;

[0148] The method further includes shortening the third dose interval after receiving one or more third doses approximately 12, 16, or 20 weeks after the preceding dose.

[0149] • Reduced from 12 weeks to 8 weeks;

[0150] • Reduced from 16 weeks to 12 weeks;

[0151] • Reduced from 16 weeks to 8 weeks

[0152] • Reduced from 20 weeks to 8 weeks

[0153] • Reduced from 20 weeks to 12 weeks, or

[0154] • Reduced from 20 weeks to 16 weeks.

[0155] This invention provides a VEGF receptor fusion protein for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, the treatment and prevention comprising administering three doses of approximately ≥8 mg of the VEGF receptor fusion protein to the eye of the subject at 4-week intervals in the form of a formulation containing approximately 114.3 mg / ml of the VEGF receptor fusion protein; wherein, after said three doses, one or more doses of the VEGF receptor fusion protein are administered at intervals extended to 12 weeks, 16 weeks, or 20 weeks.

[0156] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, the treatment and prevention comprising administering a single initial dose of about 8 mg or more of the VEGF receptor fusion protein to the eye of the subject, followed by two second doses of about 8 mg or more of the VEGF receptor fusion protein.

[0157] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and, after said dose...

[0158] a) Determine whether the subject meets at least one criterion for reducing or extending one or more intervals between VEGF receptor fusion protein doses by 2 weeks, 3 weeks, 4 weeks, or 2–4 weeks; and

[0159] b) If the determination is made, administer a higher dose of the VEGF receptor fusion protein at the reduced or extended dose interval.

[0160] The criteria for extending the interval include:

[0161] 1. BCVA loss > 5 letters;

[0162] 2. Increase in central retinal thickness (CRT) > 25 micrometers;

[0163] 3. New central fossa hemorrhage; and / or

[0164] 4. New vascularization in the central fovea

[0165] Furthermore, the criteria for reducing the interval include:

[0166] 1. BCVA loss < 5 letters,

[0167] 2. There is no liquid in the central partition;

[0168] 3. No new central foveal hemorrhage; and / or

[0169] 4. No new blood vessels formed in the central fovea.

[0170] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects who have been pretreated with one or more 2 mg doses of the VEGF receptor fusion protein. The method comprises administering a single initial dose of about 8 mg or more of the VEGF receptor fusion protein to the eye of the subject, followed by one or more administrations of a second dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more administrations of a third dose of about 8 mg or more of the VEGF receptor fusion protein.

[0171] Each second dose is administered approximately 2 to 4 weeks after the preceding dose; and each third dose is administered approximately 12 to 20 weeks after the preceding dose.

[0172] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of angiogenic eye disease in subjects in need, the treatment and prevention comprising administration to the subject's eye.

[0173] (1) A single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 8 weeks after the immediately preceding dose; or

[0174] (2) VEGF receptor fusion protein is administered once or more every 4 weeks at doses of 8 mg or more.

[0175] A VEGF receptor fusion protein for use in the treatment and prevention of angiogenic eye diseases, wherein the treatment or prevention includes providing [the protein] before each administration.

[0176] • A single-dose glass vial with a protective plastic cap and stopper contains approximately 70 microliters of an aqueous formulation containing 8 mg of VEGF receptor fusion protein;

[0177] • A 18-gauge x 1 1 / 2-inch, 5-micron filter needle, including a tip and bevel;

[0178] Packaged together; then

[0179] (1) Visually inspect the aqueous formulation in the vial, and if particles, turbidity or discoloration are visible, use another vial containing the aqueous formulation of the VEGF receptor fusion protein;

[0180] (2) Remove the protective plastic cap from the vial; and

[0181] (3) Clean the top of the vial with an alcohol wipe; then

[0182] Use aseptic techniques:

[0183] (4) Remove the 18 x 1 1 / 2 inch, 5 micron filter needle and the 1 mL syringe from their packaging;

[0184] (5) The filter needle is attached to the syringe by screwing it onto the tip of the Luer lock syringe;

[0185] (6) Push the filter needle into the center of the stopper until the needle is fully inserted into the vial and the tip touches the bottom or bottom edge of the vial;

[0186] (7) Draw all the contents of the VEGF receptor fusion protein vial into the syringe, keep the vial upright and slightly tilted, and ensure that the bevel of the filter needle is submerged in the liquid;

[0187] (8) Continue to tilt the vial during extraction, keeping the bevel of the filter needle submerged in the formulation;

[0188] (9) When the vial is emptied, pull the plunger rod back fully to completely empty the filter needle;

[0189] (10) Remove the filter needle from the syringe and dispose of the filter needle;

[0190] (11) Remove the 30 x 1 / 2 inch injection needle from its packaging and attach the injection needle to the syringe by firmly screwing the injection needle onto the tip of the Luer lock syringe;

[0191] (12) Hold the syringe with the needle facing upwards and check for air bubbles in the syringe. If air bubbles are present, gently tap the syringe with your finger until the air bubbles rise to the top; and

[0192] (13) Slowly press down the plunger so that the tip of the plunger is aligned with the 70 microliter mark on the syringe.

[0193] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein ≥8 mg of the VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising approximately 103-126 mg / ml of the VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0194] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein ≥8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising approximately 114.3 mg / ml of the VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0195] This invention provides aflibercept for use in the treatment or prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, wherein ≥8 mg of aflibercept is in an aqueous pharmaceutical formulation, wherein the aflibercept has less than about 3.5% of high molecular weight material immediately after production and purification and / or has less than or equal to about 6% of high molecular weight material after being stored at about 2°C-8°C for about 24 months.

[0196] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein ≥8 mg of the VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising:

[0197] At least approximately 100 mg / ml of VEGF receptor fusion protein;

[0198] Approximately 10mM-100mM L-arginine;

[0199] sucrose;

[0200] Histidine-based buffer solutions; and

[0201] Surfactants;

[0202] The pH of the formulation is approximately 5.0 to approximately 6.8; the VEGF receptor fusion protein has less than approximately 3.5% high molecular weight material immediately after production and purification and / or less than or equal to approximately 6% high molecular weight material after storage at approximately 2°C to 8°C for approximately 24 months.

[0203] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein ≥8 mg of the VEGF receptor fusion protein is contained in an aqueous pharmaceutical formulation comprising:

[0204] >100 mg / ml VEGF receptor fusion protein, histidine-based buffer, and L-arginine;

[0205] • 140 mg / ml aflibercept; 20 mM histidine-based buffer; 5% sucrose; 0.03% polysorbate 20; 10 mM L-arginine; pH 5.8;

[0206] • 150+15mg / ml aflibercept, 10mM phosphate buffer, 8+0.8% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20 and 50mM L-arginine, pH 5.9-6.5;

[0207] • 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;

[0208] • 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;

[0209] • 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;

[0210] >100 mg / ml aflibercept, histidine-based buffer, and L-arginine;

[0211] • >100 mg / ml aflibercept, approximately pH 5.8, wherein the formulation forms <3% HMW aggregates after incubation at 5°C for 2 months;

[0212] • Approximately 114.3 mg / mL aflibercept; 10 mM-50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, pH 5.8;

[0213] • Approximately 114.3 mg / mL aflibercept; 10 mM His / His-HCl-based buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, pH 5.8; or

[0214] • Approximately 114.3 mg / mL aflibercept; arginine hydrochloride; histidine; histidine hydrochloride hydrate; polysorbate 20; sucrose and water for injection.

[0215] • Improved best-corrected visual acuity (BCVA);

[0216] • Best corrected visual acuity (BCVA) improved by ≥5, ≥10, ≥15, or ≥20 letters;

[0217] Best corrected visual acuity (BCVA) did not decrease;

[0218] • Eliminate retinal fluid;

[0219] • According to fluorescein angiography, there was no retinal fluid leakage.

[0220] • Reduced size of choroidal neovascularization

[0221] • No intraretinal fluid (IRF) and / or subretinal fluid were found in the central region;

[0222] • The total damaged choroidal neovascularization (CNV) area is reduced;

[0223] • Loss or reduction of intraretinal fluid;

[0224] • Loss or reduction of subretinal fluid;

[0225] • Reduced central retinal thickness (CST);

[0226] • Improved quality of life related to vision (e.g., measured by the NEI-VFQ-25 total score);

[0227] • Lack of adverse events (AEs) and / or serious AEs (SAEs) occurring during the treatment period;

[0228] • ETDRS letter rating of at least 69 (approximately 20 / 40 Snellen equivalent);

[0229] • An increase of >5, >10, >15, or >20 letters in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart, or no loss during the course of treatment;

[0230] • The increase in average BCVA over a 12-week period;

[0231] • No intraretinal fluid (IRF) or subretinal fluid;

[0232] • Reduced size of choroidal neovascularization (CNV);

[0233] • The total damage CNV area decreased compared to baseline;

[0234] • Loss of IRF and / or SRF;

[0235] • Reduced central retinal thickness (CST);

[0236] • Improvement in vision-related quality of life as measured by the National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25);

[0237] • Lack of adverse events (AEs) and / or serious AEs (SAEs) occurring during the treatment period;

[0238] • Efficacy and / or safety in subjects with nAMD were similar to aflibercept administered intravitreally at approximately 2 mg every 4 weeks for the first 3 months, followed by approximately 2 mg intravitreally every 8 weeks or every 2 months, wherein efficacy was measured as an increase in BCVA and / or a decrease in central retinal thickness, and wherein safety was measured as the incidence of adverse events such as increased blood pressure, increased intraocular pressure, visual disturbances, vitreous floaters, vitreous detachment, iris neovascularization, and / or vitreous hemorrhage;

[0239] No anti-drug antibodies were detected during treatment;

[0240] • Improvement in best corrected visual acuity (BVCA) from the start of treatment (baseline) to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60;

[0241] • An increase of ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or >7 letters in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent.

[0242] • Improvement in BCVA 4 weeks after treatment initiation is approximately 2 or 3 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 3 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0243] • The improvement in BCVA at 8 weeks after the start of treatment is approximately 5 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 4 or 5 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0244] • The improvement in BCVA at 12 weeks after treatment initiation was approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0245] • Improvement in BCVA at 16 weeks after treatment initiation was approximately 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0246] • Improvement in BCVA at 20 weeks after treatment initiation was approximately 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0247] • Improvement in BCVA at 24 weeks after treatment initiation was approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0248] • Improvement in BCVA at 28 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0249] • Improvement in BCVA at 32 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0250] • Improvement in BCVA at 36 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0251] • Improvement in BCVA at 40 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0252] • Improvement in BCVA at 44 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0253] • Improvement in BCVA at 48 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0254] • Improvement in BCVA at 52 weeks after treatment initiation was approximately 7 or 8 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0255] • Improvement in BCVA at 56 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0256] • Improvement in BCVA at 60 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0257] • Improvement in BCVA at approximately 48 to 60 weeks after treatment initiation is approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0258] • Improvement in BCVA at 60 weeks after treatment initiation is approximately 5, 10, or 15 letters (ETDRS or Snellen equivalent) when using the HDq12 or HDq16 regimens.

[0259] • BCVA improves around week 8, 9, 10, 11 or 12 after treatment initiation and is maintained during the treatment regimen (within approximately ++1 or ++2 ETDRS letters or Snellen equivalents);

[0260] • The BCVA at 4 weeks after treatment initiation is approximately 63 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 63 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0261] • The BCVA at 8 weeks after treatment initiation is approximately 65 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 65 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0262] • The BCVA at 12 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0263] • The BCVA at 16 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0264] • The BCVA at 20 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0265] • The BCVA at 24 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0266] • The BCVA at 28 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0267] • The BCVA at 32 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0268] • The BCVA at 36 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0269] • The BCVA at 40 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0270] • The BCVA at 44 weeks after treatment initiation is approximately 68 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0271] • The BCVA at 48 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0272] • The BCVA at 52 weeks after treatment initiation is approximately 67 or 68 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0273] • The BCVA at 56 weeks after treatment initiation is approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0274] • The BCVA at 60 weeks after treatment initiation is approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0275] • The BCVA between approximately 48 and 60 weeks after the start of treatment is approximately 66 to approximately 72 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 to approximately 70 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0276] • The retina has no fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the central region;

[0277] • No subretinal pigment epithelial fluid;

[0278] • According to fluorescein angiography (FA), there was no fluid leakage;

[0279] • The central retinal thickness (CRT) is reduced by at least approximately 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 micrometers;

[0280] • The change in central retinal thickness 4 weeks after the start of treatment was approximately -120, -121, -122, -122.4, or -120.2 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -126, -127, -126.6, or -126.3 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen.

[0281] • Changes in central retinal thickness 8 weeks after treatment initiation were approximately -132, -133, -134, -135, -136, -136.2, or -132.8 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -139, -140, -139.5, or -139.6 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen.

[0282] • The change in central retinal thickness 12 weeks after treatment initiation was approximately -136, -137, -138, -139, -140, -141, -140.9, or -136.6 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -144, -143, or -143.5 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 protocol;

[0283] • The change in central retinal thickness 16 weeks after treatment initiation was approximately -120 or -121 or -122 or -123 or -124 or -123.4 or -120.1 micrometers (± approximately 10, 11 or 12 micrometers) when using the HDq12 protocol; or approximately -132 or -133 or -132.1 or -133.1 micrometers (± approximately 10, 11 or 12 micrometers) when using the HDq16 protocol;

[0284] • The change in central retinal thickness 20 weeks after the start of treatment is approximately -110, -111, -112, -113, -114, -113.6, or -110.9 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -115, -116, -117, -118, -115.8, or -117.7 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 protocol;

[0285] • The change in central retinal thickness 24 weeks after treatment initiation was approximately -134, -135, -136, -137, -138, -137.6, or -134.9 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -105, -106, -107, -108, -105.3, or -107.8 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 protocol;

[0286] • The change in central retinal thickness 28 weeks after the start of treatment was approximately -130, -131, -132, -133, -134, -133.7, or -130.7 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -144, -145, -146, -147, -148, -144.7, or -147.2 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 protocol;

[0287] • The change in central retinal thickness 32 weeks after treatment initiation was approximately -118, -19, -120, -121, -120.4, or -118.1 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -141, -142, -143, -144, -141.5, or -144 micrometers (± approximately 10, 11, or 12 micrometers) when using the HDq16 protocol;

[0288] • The change in central retinal thickness at 36 weeks after treatment initiation was approximately -142, -143, -144, -144.2, or -142.2 micrometers (±10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -126, -127, -128, -129, -130, -131, -126.4, or -130.5 micrometers (±10, 11, or 12 micrometers) when using the HDq16 protocol;

[0289] • Changes in central retinal thickness at 40 weeks after treatment initiation were approximately -131, -132, -133, -134, -133.8, or -131.2 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -127, -128, or -127.5 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen.

[0290] • The change in central retinal thickness at 44 weeks after treatment initiation was approximately -120, -121, -122, -123, -124, -125, -124.7, or -120.3 micrometers (±10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -143, -144, -145, or -144.8 micrometers (±10, 11, or 12 micrometers) when using the HDq16 protocol;

[0291] • The change in central retinal thickness at 48 weeks after treatment initiation was approximately -142, -143, -144, -144.4, or -142.3 micrometers (±10, 11, or 12 micrometers) when using the HDq12 protocol; or approximately -143, -144, -145, -146, -147, -148, -143.8, or -147.1 micrometers (±10, 11, or 12 micrometers) when using the HDq16 protocol;

[0292] • The change in central retinal thickness 52 weeks after treatment initiation was approximately -143.2 micrometers (±10, 11 or 12 micrometers) with the HDq12 regimen; or approximately -139.6 micrometers (±10, 11 or 12 micrometers) with the HDq16 regimen;

[0293] • The change in central retinal thickness at 56 weeks after treatment initiation was approximately -136.3 micrometers (±10, 11, or 12 micrometers) with the HDq12 regimen; or approximately -137.5 micrometers (±10, 11, or 12 micrometers) with the HDq16 regimen.

[0294] • The change in central retinal thickness 60 weeks after the start of treatment was approximately -151.8 micrometers (±10, 11 or 12 micrometers) with the HDq12 regimen; or approximately -148.8 micrometers (±10, 11 or 12 micrometers) with the HDq16 regimen;

[0295] • At week 4 after treatment initiation, the central retinal thickness was approximately 248.2 micrometers with the HDq12 protocol and approximately 244.1 micrometers with the HDq16 protocol.

[0296] • At week 8 after treatment initiation, the central retinal thickness was approximately 234.4 micrometers with the HDq12 protocol and approximately 231.2 micrometers with the HDq16 protocol.

[0297] • The central retinal thickness at week 12 after treatment initiation was approximately 229.7 micrometers with the HDq12 protocol; or approximately 226.7 micrometers with the HDq16 protocol;

[0298] • At week 16 after treatment initiation, the central retinal thickness was approximately 247.2 micrometers with the HDq12 protocol and approximately 238.6 micrometers with the HDq16 protocol.

[0299] • At week 20 after treatment initiation, the central retinal thickness was approximately 257 micrometers with the HDq12 protocol and approximately 254.9 micrometers with the HDq16 protocol.

[0300] • At week 24 after treatment initiation, the central retinal thickness was approximately 233 micrometers with the HDq12 protocol and approximately 265.4 micrometers with the HDq16 protocol.

[0301] • At week 28 after treatment initiation, the central retinal thickness was approximately 236.9 micrometers with the HDq12 protocol and approximately 226 micrometers with the HDq16 protocol;

[0302] • At week 32 after treatment initiation, the central retinal thickness was approximately 250.2 micrometers with the HDq12 protocol and approximately 229.2 micrometers with the HDq16 protocol.

[0303] • At week 36 after treatment initiation, the central retinal thickness was approximately 226.4 micrometers with the HDq12 protocol and approximately 244.3 micrometers with the HDq16 protocol.

[0304] • At week 40 after treatment initiation, the central retinal thickness was approximately 236.8 micrometers with the HDq12 protocol and approximately 243.7 micrometers with the HDq16 protocol.

[0305] • At week 44 after treatment initiation, the central retinal thickness was approximately 245.9 micrometers with the HDq12 protocol and approximately 227.7 micrometers with the HDq16 protocol.

[0306] • At week 48 after treatment initiation, the central retinal thickness was approximately 226.2 micrometers with the HDq12 protocol and approximately 226.9 micrometers with the HDq16 protocol.

[0307] • At week 52 after treatment initiation, the central retinal thickness was approximately 227.4 micrometers with the HDq12 protocol and approximately 231.1 micrometers with the HDq16 protocol.

[0308] • At week 56 after treatment initiation, the central retinal thickness was approximately 234.3 micrometers with the HDq12 protocol and approximately 233.2 micrometers with the HDq16 protocol.

[0309] • At week 60 after treatment initiation, the central retinal thickness was approximately 218.8 micrometers with the HDq12 protocol and approximately 221.9 micrometers with the HDq16 protocol.

[0310] • The CRT decreases at approximately 4, 5, 6, 7 or 8 weeks after treatment initiation, and is maintained thereafter during the treatment regimen (within approximately ±10, ±11 or ±12 micrometers).

[0311] • Approximately 4 hours after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.0409 (±0.0605) or 0 mg / L (or <0.0156 mg / L);

[0312] • Approximately 8 hours after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.05 (±3.78), 0.0973 (±0.102), or 0.0672 mg / L, respectively.

[0313] • On approximately day 2 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.11 (±2.21), 0.146 (±0.110), or 0.0903 mg / L;

[0314] • Around day 3 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.11 (±2.06), 0.137 (±0.0947), or 0.112 mg / L;

[0315] • Around day 5 after the start of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.08 (±1.86), 0.0933 (±0.0481), or 0.0854 mg / L, respectively.

[0316] • Around day 8 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.07 (±1.75), 0.0794 (±0.0413), or 0.0682 mg / L, respectively.

[0317] • Around day 15 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.04 (±1.76), 0.0435 (±0.0199), or 0.0385 mg / L;

[0318] • Around day 22 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.02 (±1.76), 0.0213 (±0.0148), or 0.0232 mg / L, respectively.

[0319] • Around day 29 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.00766 (±0.00958) or 0 mg / L (or <0.0156 mg / L);

[0320] • Approximately 4 hours after administration of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept is approximately 0.00 mg / L;

[0321] • Approximately 8 hours after administration of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept is approximately 0.00 mg / L;

[0322] • On approximately day 2 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.06 (±3.50) or 0.124 (±0.186) mg / L;

[0323] • On approximately day 3 following the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.13 (±2.07) or 0.173 (±0.155) mg / L.

[0324] • On approximately day 5 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.18 (±1.88) or 0.223 (±0.157) mg / L;

[0325] • Around day 8 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.31 (±1.56) or 0.334 (±0.135) mg / L;

[0326] • On day 15 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.37 (±1.50) or 0.393 (±0.130) mg / L.

[0327] • On approximately day 22 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.25 (±3.00) or 0.335 (±0.155) mg / L;

[0328] • On day 29 after initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.32 (±1.39) or 0.331 (±0.0953) mg / L.

[0329] • Non-inferiority of BVCA compared to intravitreal administration of aflibercept at approximately 2 mg every 4 weeks for the first 5 injections, followed by approximately 2 mg every 8 weeks or every 2 months.

[0330] • Ocular and non-ocular safety or mortality in subjects with DME was similar to that of aflibercept administered intravitreally at approximately 2 mg every 4 weeks for the first 3 or 4 or 5 injections, followed by intravitreal administration at approximately 2 mg every 8 weeks or every 2 months.

[0331] • Optimal improvement in corrected visual acuity from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48;

[0332] • From the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60, an increase in best corrected visual acuity of ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or >7 letters as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent;

[0333] • From the start of treatment until weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60, the retina showed no fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the central region; and / or

[0334] • From the start of treatment until weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56 or 60, the central retinal thickness (CRT) is reduced by at least 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150 micrometers.

[0335] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects of need, wherein:

[0336] • During the first year, the subjects were administered ≥8 mg of VEGF receptor fusion protein at one initial dose, two second doses, and three third doses.

[0337] • During the first year, the subjects were administered ≥8 mg of the VEGF receptor fusion protein, consisting of one initial dose, two second doses, and two third doses; or

[0338] • The subjects were given ≥8 mg of the VEGF receptor fusion protein once in the first year, twice in the second year, and three times in the third year, followed by two to four times in the second year.

[0339] This invention provides a VEGF receptor fusion protein for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, wherein the dose interval of ≥8 mg VEGF receptor fusion protein is adjusted (increased / maintained / decreased) based on visual and / or anatomical results.

[0340] This invention provides VEGF receptor fusion proteins for use in the treatment and prevention of neovascular age-related macular degeneration (nAMD) in subjects in need, wherein these doses of ≥8 mg of VEGF receptor fusion protein are administered according to a dosing regimen of on-demand (PRN), capped PRN, or treatment and extension (T&E).

[0341] 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 invention, the container is a vial or a pre-filled syringe, the vial being a Type I glass vial containing a nominal fill volume of approximately 0.26 mL of intravitreal injection solution. In one embodiment of the invention, the container contains a VEGF receptor fusion protein at a concentration greater than or equal to 100 mg / mL, or the container contains aflibercept at a concentration of approximately 114.3 mg / mL. In one embodiment of the invention, the instructions for use include instructions for treating DME and / or AMD with the VEGF fusion protein or aflibercept. In one embodiment of the invention, the instruction manual contains the following information: i) the container contains ≥8 mg (114.3 mg / mL) of aflibercept solution for intravitreal injection; ii) each single-dose vial provides a usable amount to deliver a single 70 μL dose containing 8 mg aflibercept to an adult patient; iii) the recommended dose is ≥8 mg aflibercept (equivalent to 70 μL of injection solution); iv) treatment with ≥8 mg aflibercept begins with one injection per month (every 4 weeks) for three consecutive doses; v) the injection interval can then be extended to every 16 or 20 weeks; vi) the treatment interval can be adjusted based on the physician’s judgment of visual and / or anatomical results; 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. Attached Figure Description

[0342] Figure 1 Summary of PULSAR clinical trials.

[0343] Figure 2 Key eligibility criteria (inclusion and exclusion criteria) for PULSAR clinical trials.

[0344] Figure 3 Dosing schedule and dosage regimen modification (DRM) standards for PULSAR clinical trials.

[0345] Figure 4 Dosage Regimen Modification (DRM) Standards for PULSAR Clinical Trials.

[0346] Figure 5 Patient management at week 48 of the PULSAR clinical trial.

[0347] Figure 6 Baseline demographic data of participants in the PULSAR clinical trial.

[0348] Figure 7 The baseline characteristics of the eyes of subjects were studied in the PULSAR clinical trial.

[0349] Figure 8 The average number of injections by week 48 in the PULSAR clinical trial.

[0350] Figure 9 *The mean change in best corrected visual acuity (BCVA) at week 48 in the PULSAR clinical trial.

[0351] The least squares mean change from baseline at week 48 is shown (see data below).

[0352]

[0353]

[0354] *This invention includes a method for achieving a specified improvement in BVCA approximately at a specified time point when receiving a specified treatment regimen for treating wet AMD.

[0355] Figure 10 The percentage of subjects who maintained the Q12 and Q16 intervals through week 48 in the PULSAR clinical trial.

[0356] Figure 11 The key secondary endpoint in the PULSAR clinical trial was the percentage of participants without retinal fluid in the central region at week 16.

[0357] Figure 12 The percentage of subjects without retinal fluid in the central region at week 48 in the PULSAR clinical trial.

[0358] Figure 13A and Figure 13B: The average change in central retinal thickness from baseline by week 48 ( Figure 13A )

[0359]

[0360] *This invention includes a method for achieving approximately a specified reduction in central retinal thickness at a specified time point when receiving a specified treatment regimen for treating wet AMD.

[0361] ; and the central retinal thickness up to week 48 ( Figure 13B (See data below).

[0362]

[0363] *This invention includes a method for achieving approximately a specified central retinal thickness at a specified time point when receiving a specified treatment regimen for treating wet AMD.

[0364] Figure 14A and 14B Serious treatment-emergent adverse events (TEAEs) in the eyes at week 48 of the PULSAR clinical trial. Figure 14A The most common ocular adverse events (AEs) by week 48 were... Figure 14B ).

[0365] Figure 15 Intraocular inflammation occurred during the 48th week of treatment in the PULSAR clinical trial.

[0366] Figure 16 The average change in intraocular pressure from baseline at week 48 of the PULSAR clinical trial.

[0367] Figure 17 The percentage of subjects who met the intraocular pressure criteria in the PULSAR clinical trial.

[0368] Figure 18 In the PULSAR clinical trial, the percentage of non-ocular serious TEAEs at week 48 was ≥0.5%.

[0369] Figure 19A and 19B Antiplatelet trial participant collaboration (APTC) events that occurred during the 48th week of treatment in the PULSAR clinical trial. Figure 19A Non-ocular safety at week 48 Figure 19B ).

[0370] Figure 20 Hypertensive events that occurred during the treatment period up to week 48 of the PULSAR clinical trial.

[0371] Figure 21Potentially clinically significant values ​​(PCSV) of blood pressure at week 48 in the PULSAR clinical trial.

[0372] Figure 22 The mean change in systolic blood pressure from baseline to week 48 in the PULSAR clinical trial is shown in the inset. The inset plot shows the mean change from baseline to week 9 and the mean baseline blood pressure.

[0373] Figure 23 The mean change in diastolic blood pressure from baseline to week 48 in the PULSAR clinical trial is shown in the inset. The inset plot illustrates the mean change from baseline to week 9 and the mean baseline blood pressure.

[0374] Figure 24 The patient died at week 48 during the PULSAR clinical trial.

[0375] Figure 25 PULSAR dosing schedule up to week 60. Dosing regimen modifications are illustrated in the illustrations.

[0376] Figure 26 Absolute BCVA and BCVA variation (ETDRS letters) from baseline to week 60. The least squares mean change from baseline is shown at week 60.

[0377] Figure 27 The proportion of patients who maintained the HDq12(8q12)- and HDq16(8q16) intervals by week 60.

[0378] Figure 28 The average number of injections by week 60 in each group.

[0379] Figure 29A , Figure 29B and Figure 29C Central retinal thickness (CRT) and its change from baseline up to week 60. (A) Central retinal thickness (µm) over time (observations - review data after ICE); (B) Mean change from baseline of CST (central retinal thickness (convertible with CRT); µm) at week 60 visit in the full analysis set, prior to ICE; (C) CST (µm) at MMRM (mixed model for repeated measures) (up to week 48) at visit in the full analysis set, compared with baseline. mean (95% CI) change.

[0380] Figure 30A , Figure 30B , Figure 30C , Figure 30D , Figure 30E , Figure 30F and Figure 30G For PULSAR, security data summary: ( Figure 30A) By week 60, ocular TEAE ≥ 2%, Figure 30B Severe TEAE of the eye at week 60, Figure 30C By week 60, non-ocular TEAE ≥2%, Figure 30D By week 60, the percentage of non-ocular severe TEAEs ≥0.5% Figure 30E Death at week 60, Figure 30F The mean change in systolic blood pressure at week 60, and ( Figure 30G Mean change in diastolic blood pressure at week 60.

[0381] Figure 31 Changes in letters 5, 10, and 15 at week 60. Observation (OC) (Data review after ICE)

[0382] Figure 32 By the week 60 visit (weeks and dose schedules shown), the percentage of subjects without retinal fluid in the central region was [percentage missing]. (LOCF - data review after ICE)

[0383] Figure 33 Structural representation of population pharmacokinetic models after aflibercept administration via IV, SC, and IVT. CMT = compartment, IV = intravenous, IVT = intravitreal, K20 = elimination rate constant of free aflibercept, K40 = adjusted elimination rate constant of bound aflibercept, K62 = absorption rate from subcutaneous reservoir compartment, K70 = elimination rate constant from tissue (platelet) compartment; QE = intercompartmental clearance of free aflibercept between the ocular ventricle and central compartment, QF1 and QF2 = intercompartmental clearance of free aflibercept, VMK24, KM = saturated Michaelis-Menten type binding of free aflibercept to VEGF; VMK27, KMK27 = saturated elimination from plasma compartment to tissue compartment (platelet), CMT 2 and CMT 4 are representative of plasma compartments and are assumed to be of equal volume.

[0384] Figure 34In the Dense PK Sub-Study (DPKS, Log-Scale), mean (±SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days at a single 2 mg and 8 mg IVT dose in nAMD or DME. LQ = Below the limit of quantitation, DME = Diabetic macular edema, DPKS = Intensive pharmacokinetic substudy, HDq12 = 8 mg aflibercept every 12 weeks after 3 initial monthly injections, HDq16 = 8 mg aflibercept every 16 weeks after 3 initial monthly injections, IVT = Intravitreal, LLOQ = Lower limit of quantitation, N = Number of participants, nAMD = Neovascular age-related macular degeneration, SD = Standard deviation adjusted bound aflibercept = 0.717 * bound aflibercept. Note: Concentrations below LLOQ (free aflibercept concentration of 0.0156 mg / L, adjusted bound aflibercept concentration of 0.0224 mg / L) are set as LLOQ / 2. Note: Data on 8 mg HD aflibercept for the first 28 days (obtained from PULSAR or PHOTON) are a combination of data from participants who received HDq12 or HDq16. One participant with an outlier free aflibercept concentration greater than 10 times the mean concentration on day 28 in PULSAR was excluded. Records after treatment in the ipsilateral eye were excluded. Data source: Drug concentration data from the week 48 database locks of PULSAR and PHOTON and the final locks of CANDELA.

[0385] Figure 35 Stratified by dose and population, observed and model-predicted plasma concentrations (mg / L) of free aflibercept and adjusted bound aflibercept within 28 days following a single IVT injection in participants with nAMD or DME in intensive PK sub-studies (DPKS). DME = diabetic macular edema, IVT = intravitreal, LLOQ = lower limit of quantitation, nAMD = neovascular age-related macular degeneration, PK = observed pharmacokinetic concentration 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 intensive PK sub-studies in PHOTON, PULSAR, and CANDELA.

[0386] Figure 36The sum of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in the plasma of the nAMD and DME combined populations. 2q8 = 2 mg aflibercept every 8 weeks after 3 initial injections at 4-week intervals; 2q12 = 2 mg aflibercept every 12 weeks after 3 initial injections at 4-week intervals; DME = diabetic macular edema; HDq12 = 8 mg aflibercept every 12 weeks after 3 initial monthly injections; HDq16 = 8 mg aflibercept every 16 weeks after 3 initial monthly injections; IVT = intravitreal; LLOQ = lower limit of quantitation; nAMD = neovascular age-related macular degeneration; 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) are set as LLOQ / 2. Data source: Drug concentration data from CANDELA, PHOTON, and PULSAR.

[0387] Figure 37 In participants with a combination of nAMD and DME, the model predicted aflibercept exposure (mg) after a single IVT injection was stratified by dosing regimen. DME = diabetic macular edema, HD = aflibercept 8 mg, IVT = intravitreal, nAMD = neovascular age-related macular degeneration, PI = predicted interval, PK = pharmacokinetics, QE = intercompartmental clearance of free aflibercept between the ocular ventricle and central compartment. The adjusted LLOQ (0.0624 μg) was set as the LLOQ of the free aflibercept concentration in plasma (i.e., 0.0156 mg / L) multiplied by the assumed volume of the study ocular ventricle in the PK model (i.e., 4 mL). Since the concentration of intraocular (free or bound) aflibercept was not measured in the clinical studies included in the population PK analysis dataset, this target was arbitrarily chosen based on the LLOQ in plasma and used as a reference for comparisons between dosing regimens to assess the effect of HD aflibercept on QE.

[0388] Figure 38In the Dense PK Sub-Study (DPKS, Log-Scale), the mean (±SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days with a single 2 mg and 8 mg IVT dose in participants with nAMD were as follows – no outliers. DME = Diabetic Macular Edema, DPKS = Dense Pharmacokinetic Sub-Study, HDq12 = 8 mg aflibercept administered every 12 weeks after 3 initial monthly injections, HDq16 = 8 mg aflibercept administered every 16 weeks after 3 initial monthly injections, IVT = Intravitreal, LLOQ = Lower Limit of Quantification, N = Number of Participants, nAMD = Neovascular Age-Related Macular Degeneration, PK = Pharmacokinetics, SD = Standard Deviation Adjusted Bound Aflibercept = 0.717 * Bound Aflibercept. Note: Concentrations below LLOQ (free aflibercept concentration of 0.0156 mg / L and adjusted bound aflibercept concentration of 0.0224 mg / L) are defined as LLOQ / 2. Note: The 8 mg data for the first 28 days (obtained from PULSAR or PHOTON) are a combination of data from participants who received HDq12 or HDq16. Data source: Drug concentrations at week 48 from PULSAR and PHOTON and final locks from CANDELA. Records after ipsilateral eye treatment were excluded.

[0389] Figure 39 In the Dense PK Sub-Study (DPKS, Log-Scale), the mean (±SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days with a single 2 mg and 8 mg IVT dose in participants with nAMD – including outliers. DME = Diabetic macular edema, DPKS = Dense Pharmacokinetic Sub-Study, HDq12 = 8 mg aflibercept every 12 weeks following 3 initial monthly injections, HDq16 = 8 mg aflibercept every 16 weeks following 3 initial monthly injections, IVT = Intravitreal, LLOQ = Lower limit of quantitation, N = Number of participants, nAMD = Neovascular age-related macular degeneration, PK = Pharmacokinetics, SD = Standard deviation adjusted bound aflibercept = 0.717 * bound aflibercept. Note: Concentrations below the lower limit of quantitation (LLOQ, free aflibercept concentration of 0.0156 mg / L, adjusted bound aflibercept concentration of 0.0224 mg / L) are defined as LLOQ / 2. Data source: Drug concentrations at week 48 of PULSAR and final lockouts of CANDELA. Data from VGFTOD-0702 are used as a reference (concentrations in the PK sub-study were subtracted from pre-dose concentrations when >LLOQ). Records after ipsilateral eye treatment were excluded.

[0390] Figure 40In the Dense PK Subset Study (DPKS, Log-Scale), the mean (±SD) concentrations (mg / L) of free and adjusted bound aflibercept over 28 days with a single 2 mg and 8 mg IVT dose in participants with DME. BLQ = below the limit of quantitation, DME = diabetic macular edema, DPKS = Dense Pharmacokinetic Set, HDq12 = 8 mg aflibercept every 12 weeks following 3 initial monthly injections, HDq16 = 8 mg aflibercept every 16 weeks following 3 initial monthly injections, IVT = intravitreal, LLOQ = lower limit of quantitation, N = number of participants, nAMD = neovascular age-related macular degeneration, SD = standard deviation. Note: Concentrations below LLOQ (free aflibercept concentration of 0.0156 mg / L and adjusted bound aflibercept concentration of 0.0224 mg / L) are defined as LLOQ / 2. Adjusted bound aflibercept = 0.717 * bound aflibercept. Note: 8 mg data for the first 28 days (obtained from PULSAR or PHOTON) are a combination of data from participants who received HDq12 or HDq16. Note: If a participant took aflibercept before starting the study drug within 12 weeks and had a baseline concentration >BLQ, that concentration was subtracted from the baseline concentration. Data source: Drug concentration data from PHOTON week 48 lock. Included drug concentration data (historical data) from VGFT-OD-0706 for reference. Records after ipsilateral eye treatment were excluded.

[0391] Figure 41 The sum of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in the plasma of participants with nAMD. 2q8 = 2 mg aflibercept every 8 weeks after 3 initial injections at 4-week intervals; 2q12 = 2 mg aflibercept every 12 weeks after 3 initial injections at 4-week intervals; DME = diabetic macular edema; HDq12 = 8 mg aflibercept every 12 weeks after 3 initial monthly injections; HDq16 = 8 mg aflibercept every 16 weeks after 3 initial monthly injections; IVT = intravitreal; LLOQ = lower limit of quantitation; nAMD = neovascular age-related macular degeneration; PK = pharmacokinetics, with 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) set as LLOQ / 2. Data from PULSAR and CANDELA.

[0392] Figure 42This is the sum of observed and model-predicted concentrations (mg / L) of free and adjusted bound aflibercept in the plasma of participants with diabetic macular edema in the PHOTON study. 2q8 = 2 mg aflibercept every 8 weeks following 3 initial injections at 4-week intervals; HDq12 = 8 mg aflibercept every 12 weeks following 3 initial injections at 4-week intervals; HDq16 = 8 mg aflibercept every 16 weeks following 3 initial injections at 4-week intervals; IVT = intravitreal; LLOQ = lower limit of quantitation. Data from PHOTON. Detailed Implementation

[0393] This invention partially provides a safe and effective high-dose aflibercept IVT injection that extends the maintenance dosing interval to more than 8 weeks, with at least similar functionality and potentially improved anatomical outcomes. This regimen demonstrated a surprisingly high level of durability in subjects, exceeding the expected durability based solely on administering more aflibercept.

[0394] EYLEA has become the standard of care for nAMD. Eylea is used for nAMD at a dose of 2 mg once monthly for three months, followed by maintenance every 8 weeks. The dosing regimen of this invention has demonstrated that a substantial percentage of subjects can maintain the 12-week and 16-week dosing intervals. In trials testing these dosing regimens, nearly 90% of subjects with diabetic macular edema were able to maintain the 16-week dosing regimen. These durability data, coupled with a safety profile consistent with EYLEA, support high-dose aflibercept as a potential new standard of care for angiogenic eye diseases such as nAMD. The data presented here demonstrate that aflibercept 8 mg dosing regimens for 12 and 16 weeks achieved high levels of improvement in visual acuity and retinal fluid anatomy measurements in subjects with nAMD, maintaining these improvements over 48 weeks. These results were obtained in subjects who rapidly initiated extended dosing intervals, the vast majority of whom did not require regimen modifications. In summary, the key data support that aflibercept 8 mg provides a longer duration of action while maintaining a similar safety profile to EYLEA.

[0395] Prior to the initiation of the HD aflibercept clinical development program, pharmacokinetic simulations using a one-chamber eye model to model the concentration-time profile of free aflibercept in the human vitreous body predicted that an 8 mg IVT dose of aflibercept would extend the dosing interval by approximately 20 days (two half-lives) compared to a 2 mg IVT dose. The HDq12 and HDq16 regimens of aflibercept demonstrated a longer duration of efficacy in the HD clinical studies than predicted. Subsequent population pharmacokinetic analyses integrating data from the CANDELA PHOTON and PULSAR Phase 3 studies indicated that the intraocular clearance of free aflibercept in HD was 34% slower than that of 2 mg IVT aflibercept administered as Eylea. This slower intraocular clearance of HD aflibercept was expected to result in a longer persistence of free aflibercept in the eye and a duration of efficacy approximately 6 weeks longer compared to 2 mg. Compared to 2 mg Eylea, the reduction in ocular clearance with HD aflibercept was greater than expected and was attributed to the "HD aflibercept effect," a highly statistically significant effect in a population pharmacokinetic model that cannot be explained simply by increasing the dose from 2 mg to 8 mg.

[0396] Population pharmacokinetic (PK) estimates suggest a median time to reach the lower limit of quantitation (LLOQ) of free aflibercept in plasma after 2 mg IVT aflibercept, compared to 3.5 weeks for 8 mg HD aflibercept. The longer duration of systemic exposure to free aflibercept (representing the movement of free aflibercept from the eye) with the HD aflibercept regimen is attributed not only to the higher dose and nonlinear systemic targeting-mediated elimination, but also to a 34% slower ocular clearance of free aflibercept. The slower ocular clearance of HD aflibercept compared to 2 mg aflibercept is projected to provide a longer 6-week duration of efficacy because, for HD aflibercept, the population pharmacokinetic estimate of reaching the amount of free aflibercept in the ocular ventricle at the end of the 8-week dosing interval occurs after 6 weeks. Exposure-response analysis estimated that the slower ocular clearance of 8 mg aflibercept could be attributed to the effect of the HD drug, resulting in a dosing regimen modification (DRM) rate that was 20.6% lower than expected if the HD drug had the same ocular clearance as 2 mg aflibercept.

[0397] General methods for protein purification are described, including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization (Coligan, et al. (2000), Current Protocols in Protein Science, Vol. 1, John Wiley & Sons, New York). Describe chemical analysis, chemical modification, post-translational modification, fusion protein production, and protein glycosylation (see, for example, Coligan et al. (2000), *The Latest Laboratory Guide to Protein Science*, Vol. 2, John Wiley & Co., New York; Ausubel et al. (2001), *Experimental Techniques in Modern Molecular Biology*, Vol. 3, John Wiley & Co., New York, pp. 16.0.5–16.22.17; Sigma-Aldrich, Co. (2001), *Products for Life Science Research*, St. Louis, Missouri, Mo., pp. 45–89; Amersham Pharmacia Biotech (2001), *BioDirectory*, Piscataway, NJ, pp. 384–391). Describe the generation, purification, and fragmentation of polyclonal and monoclonal antibodies (Coligan, et al. (2001), Current Protocols in Immunology, Vol. 1, John Wiley & Co., New York; Harlow and Lane (1999), Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Harlow and Lane, ibid.). Standard techniques for characterizing ligand / receptor interactions are available (see, for example, Cooligan, et al. (2001), Current Protocols in Immunology, Vol. 4, John Wiley & Co., New York).

[0398] Flow cytometry methods, including fluorescence activated cell sorting (FACS), are available (see, for example, Owens, et al. (1994), Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley & Son Publishing, Hoboken, NJ; Givan (2001), Flow Cytometry, 2nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003), Practical Flow Cytometry, John Wiley & Son Publishing, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, peptides, and antibodies, including nucleic acid primers and probes, and used as diagnostic reagents, are available (Molecular Probes (2003) Catalog, Molecular Probes, Inc., Eugene, Oreg.; Sigma-Aldridge (2003) Catalog, St. Louis, Missouri).

[0399] Describe standard immune system histological methods (see, for example, 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, Philadelphia, Pennsylvania, NY; Louis, et al. (2002), Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0400] "Isolated" VEGF antagonists and VEGF receptor fusion proteins (e.g., aflibercept), peptides, polynucleotides, and carriers are at least partially free of other biomolecules from the cells or cell cultures in which they are produced. Such biomolecules include nucleic acids, proteins, other VEGF antagonists and VEGF receptor fusion proteins, lipids, carbohydrates, or other materials such as cell debris and growth media. Isolated VEGF antagonists or VEGF receptor fusion proteins may further be at least partially free of expression system components, such as biomolecules from the host cell or its growth medium. Generally, the term "isolated" is not intended to mean the complete absence of such biomolecules (e.g., possible residual traces or insignificant amounts of impurities), or the absence of water, buffers, or salts, or components of pharmaceutical formulations that include VEGF antagonists or VEGF receptor fusion proteins.

[0401] "Isolated" VEGF antagonists and VEGF receptor fusion proteins (e.g., aflibercept), peptides, polynucleotides, and carriers are at least partially free of other biomolecules from the cells or cell cultures in which they are produced. Such biomolecules include nucleic acids, proteins, other VEGF antagonists and VEGF receptor fusion proteins, lipids, carbohydrates, or other materials such as cell debris and growth media. Isolated VEGF antagonists or VEGF receptor fusion proteins may further be at least partially free of expression system components, such as biomolecules from the host cell or its growth medium. Generally, the term "isolated" is not intended to mean the complete absence of such biomolecules (e.g., possible residual traces or insignificant amounts of impurities), or the absence of water, buffers, or salts, or components of pharmaceutical formulations that include VEGF antagonists or VEGF receptor fusion proteins.

[0402] The dosages mentioned herein (e.g., 8 mg) cover embodiments including a specified amount ± 10%, such as 8 mg (± 0.8 mg). The concentrations or amounts of excipients mentioned herein also cover embodiments including a specified amount ± 10%.

[0403] The terms "subject" and "patient" are used interchangeably herein. A subject or patient is a mammal, such as a human, mouse, rabbit, monkey, or non-human primate, preferably a human. It can be said that a subject or patient "suffers from" angiogenic eye disease, such as nAMD. Such a subject or patient has the disease in one or both eyes. In one embodiment of the invention, the subject or patient (preferably a human) has one or more of the following characteristics (currently or in the past):

[0404] 1. ≥50 years old, such as 61, 62, 63, 74 or 75 years old;

[0405] 2. Having active subfoveal CNVs secondary to nAMD, for example, with a total area > 50% of the total lesion area, such as including lesions near the fovea that affect the fovea of ​​the eye;

[0406] 3. Research on early treatment of diabetic retinopathy with best-corrected visual acuity (BCVA)

[0407] (ETDRS) letter ratings are approximately 78 to 24, 73-78, <73, 58, 59, 60, 61, or 62 (Snellen equivalents are 20 / 40, 20 / 63, 20 / 50, 20 / 32, or 20 / 320), for example due to DME or nAMD;

[0408] 4. Central retinal thickness ≥300 μm or ≥320 μm or approximately 367, 368, 369, 370, 450, 451, 452, 453, 454 or 455 μm; or and / or central involvement of DME in the eye, with CRT ≥300 μm (or ≥320 μm as determined by Spectralis).

[0409] 5. The total damaged area is approximately 6 or 7 mm. 2 For example, the damage types are predominantly occult or classical, or have the lowest classical type.

[0410] 6. DRSS score is better than or equal to 43, 47 or worse;

[0411] 7. Type 1 or type 2 diabetes; and / or

[0412] 8. Intraretinal fluid and / or subretinal fluid are present in the central region of the eye as seen on an OCT scan;

[0413] 9. Body mass index (BMI) of approximately 27 or 28 or higher; and / or

[0414] 10. Hypertension; and / or

[0415] 11. History of any one or more of the following: cataracts, dry age-related macular degeneration, vitreous detachment, dry eye, nuclear cataracts, glaucoma, astigmatism, presbyopia, choroidal neovascularization, open-angle glaucoma, hyperopia, macular degeneration, myopia, posterior capsule opacification, epiretinal membrane, retinal degeneration, skin laxity, retinal hypertension, drusen, iridodialysis syndrome, vitreous floaters, cortical cataracts, ptosis, retinal tear, vitreous diseases, pterygium, vitreous degeneration, eye strain, cataract surgery, intraocular lens implantation, lens capsulotomy, blepharoplasty, keratomileusis, laser therapy, tumors (benign, malignant, and unexplained (including cysts and polyps)) or facial moles.

[0416] And / or,

[0417] It has or lacks any one or more of the following characteristics:

[0418] 1. Subretinal hemorrhage in the eye with a total damaged area ≥50%;

[0419] 2. Intraocular pressure ≥25 mmHg;

[0420] 3. Signs of infectious blepharitis, keratitis, scleritis, or conjunctivitis in the eyes;

[0421] 4. Any intraocular inflammation and / or eye infection;

[0422] 5. A history of stage II or higher macular hole in the eye;

[0423] 6. Examine the eye for current iris neovascularization, vitreous hemorrhage, or tractional retina visible during screening and evaluation.

[0424] Detachment;

[0425] 7. Blood pressure loss of control (defined as systolic blood pressure >140 mmHg or diastolic blood pressure >90 mmHg);

[0426] The change in BP measurements exceeded 10% in 8.3 measurements;

[0427] 9. Cerebrovascular accident / transient ischemic attack or myocardial infarction /

[0428] History of acute coronary syndrome;

[0429] 10. History of kidney failure, dialysis, or kidney transplantation;

[0430] 11. Known to be sensitive to aflibercept;

[0431] 12. Pregnant or breastfeeding women;

[0432] 13. Those with fertility potential* who are unwilling to undergo high-efficiency treatment.

[0433] Women who use contraception;

[0434] 14. Any intraocular surgery within 12 weeks (84 days); and / or

[0435] 15. History of corneal transplantation or corneal dystrophy.

[0436] Therefore, the present invention includes a method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject of need.

[0437] 1. Age ≥ 50 years old;

[0438] 2. It has an active central recess CNV derived from nAMD;

[0439] 3. Study on Early Treatment of Diabetic Retinopathy Based on Best Corrected Visual Acuity (BCVA)

[0440] The (ETDRS) letter score is approximately 78 to 24;

[0441] 4. The central retinal thickness is ≥300 micrometers or ≥320 micrometers;

[0442] 5. The damaged area is approximately 6 or 7 mm. 2 ;

[0443] 6. DRSS score of 43 or higher, 47 or lower; and / or

[0444] 7. Have type 1 or type 2 diabetes;

[0445] and / or

[0446] 1. No subretinal hemorrhage covering ≥50% of the total damaged area in the eye;

[0447] 2. No intraocular pressure ≥25 mmHg in the eye;

[0448] 3. The eyes show no signs of infectious blepharitis, keratitis, scleritis, or conjunctivitis;

[0449] 4. There is no intraocular inflammation and / or eye infection in the eyes;

[0450] 5. No history of stage II or higher macular hole in the eye;

[0451] 6. No visible neovascularization of the iris, vitreous hemorrhage, or tractional retinal detachment is observed in the eye;

[0452] 7. There are no uncontrolled BPs;

[0453] 8. No change exceeding 10% in the three BP measurements;

[0454] 9. No history of cerebrovascular accident / transient ischemic attack or myocardial infarction / acute coronary syndrome;

[0455] 10. No history of kidney failure, dialysis, and / or kidney transplantation;

[0456] 11. No known sensitivity to aflibercept;

[0457] 12. Women who are not pregnant or breastfeeding;

[0458] 13. Lacking the ability to reproduce and unwilling to undergo efficient [fertilization / reproduction].

[0459] Women who use contraception;

[0460] No intraocular surgery was performed within 14.12 weeks (84 days); and / or

[0461] 15. No history of corneal transplantation or corneal dystrophy in the eye;

[0462] The method involves administering a single initial dose of about 8 mg or more of VEGF receptor fusion protein (preferably aflibercept) to the eye of a subject, followed by one or more second doses of about 8 mg or more of VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediate preceding dose; and wherein each third dose is administered about 12 to 20 weeks after the immediate preceding dose.

[0463] VEGF antagonists

[0464] This invention includes methods for treating or preventing angiogenic eye diseases using VEGF antagonists. VEGF antagonists include molecules that interfere with the interaction between VEGF and its natural VEGF receptor, such as molecules that bind to VEGF or its receptor and prevent or block the interaction between VEGF and its 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 preferably used in conjunction with the methods described herein, the scope of this invention includes methods in which any of the VEGF antagonists described herein (e.g., scFv, DARPin, anti-VEGF antibodies) are used instead of such fusion proteins.

[0465] For the purposes of this document, a “VEGF receptor fusion protein” refers to a molecule containing one or more VEGF receptors or their domains, which fuses with another polypeptide to interfere with the interaction between VEGF and the native VEGF receptor, for example, by association of two such fusion polypeptides to form a homodimer or other multimer. Such VEGF receptor fusion proteins may be referred to as “VEGF-Trap” or “VEGF Trap”. VEGF receptor fusion proteins falling within the context of this disclosure as defined herein include chimeric polypeptides containing two or more immunoglobulin (Ig)-like domains of the VEGF receptor, such as VEGFR1 (also known as Flt1) and / or VEGFR2 (also known as Flk1 or KDR), and may also contain multimerizing domains (e.g., Fc domains).

[0466] An exemplary VEGF receptor fusion protein is a molecule called 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 therein.

[0467] VEGF1R2-FcΔC1(a) contains three components:

[0468] (1) VEGFR1 component, which contains amino acids 27 to 129 of SEQ ID NO:2;

[0469] (2) The VEGFR2 component, comprising amino acids 130 to 231 of SEQ ID NO:2; and

[0470] (3) The polymerized component ("FcΔC1(a)") contains 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 by reference for all purposes). Note that amino acids 1 to 26 of SEQ ID NO:2 are the signal sequence.

[0471] If the polymerized component (MC) of the VEGF receptor fusion protein originates from the Fc domain of IgG (e.g., IgG1), then the amino acid composition of the MC must be no less than amino acids 232 to 457 of SEQ ID NO:2. Therefore, the IgG component of the MC cannot be truncated to less than 226 amino acids.

[0472] In one embodiment of the invention, the VEGF receptor fusion protein comprises amino acids 27-458 or 27-457 of SEQ ID NO:2 (e.g., in the form of a homodimer).

[0473] atggtcagctactgggacaccggggtcctgctgtgcgcgctgctcagctgtctgcttctcacaggatctagttccgg

[0474] aagtgataccggtagacctttcgtagagatgtacagtgaaatccccgaaattatacacatgactgaaggaagggag

[0475] ctcgtcattccctgccgggttacgtcacctaacatcactgttatactttaaaaaagtttccacttgacactttgatccctgat

[0476] ggaaaacgcataatctgggacagtagaaagggcttcatcatatcaaatgcaacgtacaaagaaatagggcttctg

[0477] acctgtgaagcaacagtcaatgggcatttgtataagacaaactatctcacacatcgacaaaccaatacaatcataga

[0478] tgtggttctgagtccgtctcatggaattgaactatctgttggagaaaagcttgtcttaaattgtacagcaagaactgaa

[0479] ctaaatgtggggattgacttcaactgggaatacccttcttcgaagcatcagcataagaaacttgtaaaccgagacct

[0480] aaaaacccagtctgggagtgagatgaagaaatttttgagcaccttaactatagatggtgtaacccggagtgaccaa

[0481] ggattgtacacctgtgcagcatccagtgggctgatgaccaagaagaacagcacatttgtcagggtccatgaaaag

[0482] gacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttcccccca

[0483] aaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaaga

[0484] ccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggag

[0485] cagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagta

[0486] caagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccc

[0487] cgagaaccacaggtgtacaccctgcccccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcct

[0488] ggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaag

[0489] accacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtgg

[0490] cagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctcc

[0491] ctgtctccgggtaaatga

[0492] (SEQ ID NO:1)

[0493] MVSYWDTGVLLCALLSCLLLTGSSSG SDTGRPFVEMYSEIPEIIHMTEGR

[0494] ELVIPCRVTS

[0495] PNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNG

[0496] HLYKTNYLT

[0497] HRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSK

[0498] HQHKKLVNRD

[0499] LKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVH

[0500] EKDKTHTCPPC

[0501] PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY

[0502] VDGVEVHNAKT

[0503] KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS

[0504] KAKGQPREPQVY

[0505] TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL

[0506] DSDGSFFLYSK

[0507] LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0508] (SEQ ID NO:2)

[0509] In embodiments of the present invention, the VEGF receptor fusion protein comprises

[0510] (1) Immunoglobulin-like (Ig) domain 2 of the first VEGF receptor (e.g., VEGFR1), and

[0511] (2) Ig domain 3 of the second VEGF receptor (e.g., VEGFR2),

[0512] (3) and optionally, further comprising the Ig domain 4 (e.g., VEGFR2) of a second VEGF receptor and

[0513] (4) Polymerized components (e.g., the Fc domain of IgG, including the hinge, CH2 and CH3 domains).

[0514] For example, in an embodiment of the present invention, the VEGF receptor fusion protein has the following arrangement of domains:

[0515] • [VEGFR1 Ig domain 2]-[VEGFR2 Ig domain 3]-[MC] (e.g., its homodimer) or

[0516] • [VEGFR1 Ig domain 2]-[VEGFR2 Ig domain 3]-[VEGFR2 Ig domain 4]-[MC] (e.g., its homodimer).

[0517] Note that this disclosure also includes, within its scope, high-concentration formulations, including VEGF-binding molecules or anti-VEGF antibodies or their antigen-binding fragments or their biopolymer conjugates (e.g., KSI-301) (but not VEGF receptor fusion proteins), for example...

[0518] • Bevacizumab (e.g., at a concentration of approximately 80-90 or 88 mg / ml),

[0519] ranitidine (e.g., at a concentration of approximately 20-40 mg / ml, such as 21-35, 21 or 35 mg / ml),

[0520] • Anti-VEGF aptamers, such as piperatanib (e.g., piperatanib sodium),

[0521] • Single chain (e.g., V) L -V H Anti-VEGF antibodies, such as bloxicillin (e.g., concentrations of approximately 200-400 mg / ml or 200, 210, 400, or 420 mg / ml),

[0522] • Anti-VEGF DARPin, such as Abicipar Pegol DARPin (e.g., at concentrations of approximately 70-140, 70, or 140 mg / ml), or

[0523] • Bispecific anti-VEGF antibodies, such as those that also bind to ANG2, such as RG7716 (farecimab) (e.g., concentrations of approximately 100-400, 100, 105, 400, or 420 mg / ml).

[0524] To minimize the reproducibility of the embodiments discussed herein, the scope of the invention is intended to include embodiments in which any formulation discussed herein comprises any concentration of anti-VEGF antibody or antibody fragment or other VEGF-binding molecule discussed herein (e.g., replaced by anti-VEGF DARPin) instead of the VEGF receptor fusion protein. For example, the invention includes formulations containing 35 or 80 mg / ml ranizumab, buffer, heat stabilizer, viscosity reducer, and surfactant.

[0525] DARPin is a designed ankyrin repeat protein. DARPin typically contains three to four tightly packed repeat sequences of approximately 33 amino acid residues, each repeat containing one β-turn and two antiparallel α-helices. This rigid framework provides the protein with stability while enabling the presentation of variable regions, typically six amino acid residues per repeat, for target recognition.

[0526] "Anti-VEGF" antibodies or antigen-binding fragments of antibodies refer to antibodies or fragments that specifically bind to VEGF.

[0527] Exemplary VEGF receptor fusion proteins include aflibercept ( Regeneron Pharmaceuticals, Inc.) or Conbercept (marketed by Chengdu Kanghong Biotechnology Co., Ltd.). See International Patent Application Publication Nos. WO2005 / 121176 or WO2007 / 112675. The terms “aflibercept” and “Conbercept” include its biosimilars. A biosimilar version of the reference product (e.g., aflibercept) generally refers to a product containing the same amino acid sequence, but includes products biosimilar under the U.S. Biologics Price Competition and Innovation Act.

[0528] The present invention also includes embodiments in addition to VEGF antagonists, including administration of one or more other therapeutic agents, such as administration (in one or more doses) of a second VEGF antagonist, antibiotics, anesthetics (e.g., local anesthetics), nonsteroidal anti-inflammatory drugs (NSAIDs), steroids (e.g., corticosteroids, dexamethasone), triamcinolone (TA), methotrexate, rapamycin, antitumor necrosis factor α drugs (e.g., infliximab), daklizumab, and / or complement component (e.g., C3 or C5) inhibitors.

[0529] Drug formulation

[0530] This invention includes a method of administering a VEGF antagonist to the eye of a subject in a pharmaceutical formulation. The pharmaceutical formulation includes a VEGF antagonist and a pharmaceutically acceptable carrier. Other agents may be incorporated into the pharmaceutical formulation to provide improved transfer, delivery, tolerability, etc. The term "pharmaceuticalally acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopeia or other recognized pharmacopoeia for animal (more specifically, for human) use. The term "carrier" refers to a diluent, adjuvant, excipient, or medium administered with the VEGF antagonist. Numerous suitable formulations can be found in all formularies known to pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th edition, Mack Publishing Company, Easton, Pa., 1975), e.g., Chapter 87 by Blaug, Seymour.

[0531] The pharmaceutical formulation used in the method of the present invention can be "high concentration". High-concentration pharmaceutical formulations of the present invention include VEGF antagonists, such as VEGF receptor fusion proteins, with concentrations 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. "High concentration" can refer to formulations comprising VEGF antagonists with concentrations from about 140 mg / ml to about 160 mg / ml, at least about 140 mg / ml but less than 160 mg / ml, from about 41 mg / ml to about 275 mg / ml, from about 70 mg / ml to about 75 mg / ml, or from about 80 mg / ml to about 250 mg / ml. In some respects, the concentration of the VEGF antagonist in the formulation is approximately any of the following: 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 ; 59mg / ml; 60mg / ml; 61mg / ml; 62mg / ml; 63mg / ml; 64mg / ml; 65mg / ml; 66mg / ml; 67mg / ml; 68mg / ml; 69mg / ml; 70mg / ml; 71mg / ml; 72mg / ml; 73mg / ml; 74mg / ml; 75mg / 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; 91 mg / ml; 92mg / ml; 93mg / ml; 94mg / ml; 95mg / ml; 96mg / ml; 97mg / ml; 98mg / ml; 99mg / ml; 100mg / ml; 101mg / ml ;102mg / ml; 103mg / ml; 104mg / ml; 105mg / ml; 106mg / ml; 107mg / ml; 108mg / ml; 109mg / ml; 110mg / ml; 111m g / 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; 225mg / ml; 226mg / ml; 227mg / ml; 228mg / ml; 229mg / ml; 230mg / ml; 231mg / m l; 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; 251mg / 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 / ml; 269mg / ml; 270mg / ml; 271mg / ml; 272mg / ml; 273mg / ml; 274mg / ml; or 275mg / ml. Other VEGF antagonist concentrations are considered in this study, provided that the concentration is effective according to the embodiments described herein.

[0532] In embodiments of the invention, the concentration of the pharmaceutical formulation used in the method of the invention is about 4, 6, 8, 10, 12, 14, 16, 18, or 20 mg of VEGF receptor fusion protein (e.g., aflibercept), or an amount of such protein of any acceptable dose discussed herein, such as about 100 μl or less, about 75 μl or less, or about 70 μl or less, for example, 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; 68 μl; 68 μl; 69 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 ... μl; 69μl; 70μl; 71μl; 72μl; 73μl; 74μl; 75μl; 76μl; 77μl; 78μl; 79μl; 80μl; 81μl; 82μl; 83μl; 84μl ; 85μl; 86μl; 87μl; 88μl; 89μl; 90μl; 91μl; 92μl; 93μl; 94μl; 95μl; 96μl; 97μl; 98μl; 99μl; or 100μl.

[0533] This invention includes methods of using any of the formulations described under “Exemplary Formulations” (as discussed herein), but wherein the concentration of VEGF receptor fusion protein (e.g., aflibercept) is replaced by the concentration described in this section (“VEGF Receptor Fusion Proteins and Other VEGF Inhibitors”).

[0534] Buffer solutions used in pharmaceutical formulations herein that can be employed in the methods of this invention refer to solutions that resist pH changes by using acid-base conjugates. Buffer solutions are capable of maintaining a pH in the range of about 5.0 to about 6.8, more typically from about 5.8 to about 6.5, and most typically from about 6.0 to about 6.5. In some cases, the pH of the formulations of this 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 buffer solutions included in formulations herein include histidine-based buffer solutions, such as histidine with histidine hydrochloride or histidine acetate. The buffer solution included in the formulations herein may alternatively be a phosphate-based buffer (e.g., containing sodium phosphate), an acetate-based buffer (e.g., containing sodium acetate or acetic acid), or a citrate-based buffer (e.g., containing sodium citrate or citric acid). It is also recognized that the buffer solution may be a mixture of the above substances, provided that the buffer solution functions as a buffer for the formulation within the pH range described above. In some cases, the buffer solution is from about 5 mM to about 25 mM, or more typically, from about 5 mM to about 15 mM. The buffer solution may be about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, or about 25 mM.

[0535] In an embodiment of the present invention, histidine and histidine hydrochloride are used to prepare a histidine-based buffer solution.

[0536] As used herein, surfactants refer to components that protect high concentrations of VEGF antagonists from various surface and interface-induced stresses, such as VEGF receptor fusion proteins. Therefore, surfactants can be used to limit or minimize VEGF receptor fusion protein aggregation and promote protein dissolution. Suitable surfactants described herein are shown to be nonionic and may include surfactants having a polyoxyethylene moiety. Exemplary such surfactants include: polysorbate 20, polysorbate 80, poloxamer 188, polyethylene glycol 3350, and mixtures thereof. The surfactant in the formulation may be from about 0.02% to about 0.1% by weight / volume (w / v), and more typically, from 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).

[0537] Heat stabilizers used in pharmaceutical formulations according to the methods described herein refer to components that provide thermal stability against the thermal denaturation of VEGF antagonists (e.g., VEGF receptor fusion proteins) and prevent loss of efficacy or activity of the VEGF receptor fusion protein. Suitable heat stabilizers include sugars, and may be sucrose, trehalose, sorbitol, or mannitol, or may be amino acids such as L-proline, L-arginine (e.g., L-arginine monohydrochloride), or taurine. Furthermore, heat stabilizers may also include substituted acrylamides or propanesulfonic acids, or may be compounds such as glycerol.

[0538] In some cases, the pharmaceutical formulations used in the methods described herein include sugars and taurine, sugars and amino acids, sugars and propanesulfonic acid, sugars and taurine, glycerol and taurine, glycerol and propanesulfonic acid, amino acids and taurine, or amino acids and propanesulfonic acid. Additionally, formulations may include sugars, taurine and propanesulfonic acid, glycerol, taurine and propanesulfonic acid, and L-proline, taurine and propanesulfonic acid.

[0539] The embodiments described herein may have heat stabilizers present individually, 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 in combination of the above total concentrations. The concentration of the heat stabilizer in the formulation may be 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).

[0540] Regarding taurine and propanesulfonic acid, in embodiments of the invention, these heat stabilizers may be present in the formulation in amounts of about 25 mM to about 100 mM, more typically about 50 mM to about 75 mM (compared to other heat stabilizers).

[0541] Viscosity reducers are commonly used to reduce or prevent protein aggregation. Viscosity reducers included herein include sodium chloride, magnesium chloride, D- or L-arginine (e.g., L-arginine monohydrochloride), lysine, or mixtures thereof. When present herein, viscosity reducers may be present in amounts from about 10 mM to about 100 mM, more typically from about 30 mM to about 75 mM, and even more typically from about 40 mM to about 70 mM. In some cases, viscosity reducers are present in amounts of 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.

[0542] Pharmaceutical formulations used in the methods described herein may also have pharmaceutically acceptable viscosities for ocular administration, such as intravitreal injection. Viscosity is generally a measure of the resistance of a fluid to deformation under shear or tensile stress (typically measured using techniques known in the art, such as a viscometer or rheometer). Typical viscosities of formulations used in the methods described herein are from about 5.0 cP (centipoise) to about 15 cP, from about 11 cP to about 14 cP, from about 12 cP to about 15 cP, or from about 11 cP to about 12 cP. Therefore, the viscosity of the formulations described herein can be approximately 5.0 cP, approximately 6.0, approximately 7.1 cP, approximately 7.2 cP, approximately 7.3 cP, approximately 7.4 cP, approximately 7.5 cP, approximately 7.6 cP, approximately 10 cP, approximately 10.5 cP, approximately 11.0 cP, approximately 11.5 cP, approximately 12.0 cP, approximately 12.5 cP, approximately 13.0 cP, approximately 13.5 cP, approximately 14.0 cP, approximately 14.5 cP, or approximately 15.0 cP (e.g., when measured at 20°C).

[0543] The various embodiments described herein do not require the inclusion of inorganic salts or other viscosity reducers to maintain these highly useful viscosities. Typically, high-concentration protein solutions require viscosity reducers to prevent protein aggregation and higher viscosity, which makes formulations difficult to inject intravitreal and reduces the potency of VEGF receptor fusion proteins. Therefore, the embodiments described herein include methods using formulations with substantially no or no added sodium chloride (NaCl), magnesium chloride (MgCl2), D- or L-arginine (e.g., D- or L-arginine hydrochloride), lysine, or other viscosity reducers.

[0544] Osmoregulation is a key property of injectable drug formulations used in the methods of this invention. It is desirable that the product conforms to physiological osmotic conditions. Furthermore, osmoregulation provides confirmation of the soluble content in the solution. In embodiments of this invention, the molar osmoregulation of the formulation used in the methods of this invention is less than or equal to about 506 mmol / Kg or from about 250 to about 506 mmol / Kg, for example, 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 embodiments of this invention, the osmotic pressure is below about 250 mmol / Kg.

[0545] Exemplary pharmaceutical formulations for use in the methods of the present invention include the following:

[0546] Preparation 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 to 6.2.

[0547] Preparation 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 to 6.2.

[0548] Preparation 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 to 6.2.

[0549] Preparation 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.

[0550] Preparation E: 80 mg / ml aflibercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0551] Preparation 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 to 6.2.

[0552] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0553] Formulation H: 80 mg / ml aflibercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0554] Formulation I: 80 mg / ml aflibercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0555] 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 to 6.2, and optionally, particularly, excluding viscosity reducers.

[0556] Formulation K: 80 mg / ml aflibercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0557] Formulation L: 80 mg / ml aflibercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0558] Preparation 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 to 6.2.

[0559] Preparation N: 150 mg / ml aflibercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0560] Preparation 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 to 6.2.

[0561] Preparation 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.

[0562] Preparation Q: 150 mg / ml aflibercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0563] Preparation 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 to 6.2.

[0564] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0565] Formulation T: 150 mg / ml aflibercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2 (e.g. 6.2), and optionally, specifically, excluding viscosity reducers.

[0566] Formulation U: 150 mg / ml aflibercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0567] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0568] The formulation W consists of 150 mg / ml aflibercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, at pH 5.8 to 6.2, and optionally, specifically, excludes viscosity reducers.

[0569] Formulation X: 150 mg / ml aflibercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0570] 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 to 6.2.

[0571] Preparation Z: 80 mg / ml Conbercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0572] Preparation 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 to 6.2.

[0573] Preparation 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.

[0574] Preparation CC: 80 mg / ml Conbercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0575] Preparation 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 to 6.2.

[0576] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0577] Formulation FF: 80 mg / ml conbercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0578] Formulation GG: 80 mg / ml conbercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0579] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0580] Formulation II: 80 mg / ml Conbercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0581] Formulation JJ: 80 mg / ml conbercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0582] Preparation 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 to 6.2.

[0583] Preparation LL: 150 mg / ml Conbercept, 10 mM phosphate-buffered saline, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20 and 40 mM sodium chloride, pH 5.8 to 6.2.

[0584] Preparation 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 to 6.2.

[0585] Preparation 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.

[0586] Preparation OO: 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 to 6.2.

[0587] Preparation 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 to 6.2.

[0588] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0589] The formulation RR consists of 150 mg / ml conbercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, at pH 5.8 to 6.2, and optionally, specifically, excludes viscosity reducers.

[0590] The formulation SS consists of 150 mg / ml conbercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, at pH 5.8 to 6.2, and optionally, specifically, excludes viscosity reducers.

[0591] 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 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0592] The formulation UU consists of 150 mg / ml conbercept, 10 mM phosphate-buffered saline, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, at pH 5.8 to 6.2, and optionally, specifically, excludes viscosity reducers.

[0593] Formulation VV: 150 mg / ml Conbercept, 10 mM citrate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally, specifically, excluding viscosity reducers.

[0594] The formulation consists of 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, at pH 5.8.

[0595] 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.

[0596] 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.

[0597] 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.

[0598] 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.

[0599] The formulation BBB consists of 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, at pH 5.8.

[0600] Preparation 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.

[0601] The formulation consists of 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, at pH 5.8.

[0602] Formula 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, and optionally without heat stabilizers, pH 5.8.

[0603] Formula 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.

[0604] 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

[0605] Formulation HHH: 140 mg / ml VEGF receptor fusion protein formulation (e.g., aflibercept); 20 mM histidine, pH 5.8; 5% sucrose; 0.03% polysorbate 20; 50 mM sodium thiocyanate

[0606] 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

[0607] 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

[0608] 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.

[0609] 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

[0610] Formulation MMM: 140 mg / ml VEGF receptor fusion protein formulation (e.g., aflibercept); 20 mM histidine, pH 5.8; 5% sucrose; 0.03% polysorbate 20; 50 mM sodium aspartate

[0611] Formulation NNN: 140 mg / ml VEGF receptor fusion protein formulation (e.g., aflibercept); 20 mM histidine, pH 5.8; 5% sucrose; 0.03% polysorbate 20; 50 mM sodium glutamate.

[0612] Formulation OOO: 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).

[0613] 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).

[0614] Formulation QQQ: 140 mg / ml VEGF receptor fusion protein formulation (e.g., aflibercept); 20 mM histidine, pH 5.8; 5% sucrose; 0.03% polysorbate 20; 50 mM sodium aspartate; 50 mM arginine (e.g., arginine hydrochloride).

[0615] 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 glutamate; 50 mM arginine (e.g., arginine hydrochloride).

[0616] 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).

[0617] 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 L-arginine (L-arginine hydrochloride)

[0618] Formulation UUU: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0619] Formulation VVV: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0620] Formulation: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0621] Formulation XXX: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0622] Formulation YYY: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0623] Formulation ZZZ: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0624] 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

[0625] 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

[0626] Preparation 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

[0627] The formulation consists of 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), at pH 5.8.

[0628] 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-hydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6–6.0 or 5.5–6.1).

[0629] The formulation consists of 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 (L-arginine monohydrochloride) (e.g., ±5 mM), at pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0630] Formulation GGGG: 114.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., 103-126 mg / ml), 10 mM histidine-based buffer (e.g., including 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).

[0631] 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 hydrochloride-hydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6–6.0 or 5.5–6.1).

[0632] Formulation IIII: 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 hydrochloride-hydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6–6.0 or 5.5–6.1).

[0633] Formula 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 (L-arginine hydrochloride), pH 6.2 (e.g., 6.0-6.4 or 5.9-6.5).

[0634] 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., arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6–6.0 or 5.5–6.1);

[0635] See International Patent Application Publication No. WO2019 / 217927.

[0636] In one embodiment of the invention, ≥8 mg of VEGF receptor fusion protein (preferably aflibercept) is administered in an aqueous pharmaceutical formulation comprising: the VEGF receptor fusion protein comprising two polypeptides, each polypeptide comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a polymerized component (e.g., containing 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 HCl); and about 0.03% surfactant; wherein the pH of the formulation is about 5.0 to about 6.8 (e.g., 5.8 to 6.5, e.g., 5.8). Preferably, the formulation is suitable for intravitreal administration. Other components that may be included are sodium sulfate, sodium thiocyanate, glycine, NaCl, sodium aspartate, and / or monosodium glutamate. In one embodiment of the invention, the concentration of the VEGF receptor fusion protein is: 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) a molar osmotic pressure of about 299 to about 506 mmol / Kg; and / or (ii) a viscosity of about 6-15 cP at 20°C. The surfactant may be a nonionic surfactant, such as polysorbate 20, polysorbate 80, poloxamer 188, polyethylene glycol 3350, or mixtures thereof. The concentration of the histidine-based buffer may be from about 10 mM to 20 mM. In one embodiment of the invention, the VEGF receptor fusion protein has less than about 3.5% high molecular weight material immediately after production and purification and / or less than or equal to about 6% high molecular weight material after being stored at about 2°C to 8°C for about 24 months.

[0637] In one embodiment of the invention, ≥8 mg of VEGF receptor fusion protein is administered in an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of VEGF receptor fusion protein, the fusion protein comprising two polypeptides, each polypeptide comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a polymerized component; about 10 mM-100 mM L-arginine; sucrose; a histidine-based buffer; and a surfactant; wherein the pH of the formulation is about 5.0 to about 6.8; wherein the VEGF receptor fusion protein has less than about 3.5% high molecular weight material immediately after production and purification and / or less than or equal to about 6% high molecular weight material after storage at about 2°C-8°C for about 24 months.

[0638] In one embodiment of the present invention, the aqueous formulation comprises:

[0639] • ≥100 mg / ml VEGF receptor fusion protein (e.g., aflibercept), histidine-based buffer, and L-arginine;

[0640] • Approximately 140 mg / ml aflibercept; 20 mM histidine-based buffer; 5% sucrose; 0.03% polysorbate 20; 10 mM L-arginine; pH 5.8;

[0641] • Approximately 150±15 mg / ml aflibercept, 10 mM phosphate-buffered saline, 8±0.8% (w / v) sucrose, 0.02-0.04% (w / v) polysorbate 20 and 50 mM L-arginine, pH 5.9-6.5;

[0642] • 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;

[0643] • 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;

[0644] • 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;

[0645] • ≥100 mg / ml aflibercept, histidine-based buffer, and L-arginine;

[0646] • ≥100 mg / ml aflibercept, pH 5.8, wherein the formulation forms <3% HMW aggregates after incubation at 5°C for 2 months;

[0647] • Approximately 114.3 mg / mL aflibercept; 10 mM-50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, pH 5.8;

[0648] or

[0649] • Approximately 114.3 mg / mL aflibercept; 10 mM His / His-HCl based buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, pH 5.8.

[0650] In one embodiment of the invention, ≥8 mg of VEGF receptor fusion protein is administered in an aqueous pharmaceutical formulation containing...

[0651] 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);

[0652] Heat stabilizers, which are sugars, amino acids, sucrose, mannitol, sorbitol, trehalose, L-proline, glycine, glycerol, taurine, or propanesulfonic acid (e.g., about 2% (w / v) to about 10% (w / v), such as 5% (w / v));

[0653] Buffer solutions, which are histidine-based buffer solutions, phosphate-based buffer solutions, acetate-based buffer solutions (e.g., at a concentration of about 5-25 mM, such as 10 mM or 20 mM); or citrate-based buffer solutions;

[0654] Nonionic surfactants, such as polysorbate 20, polysorbate 80, poloxamer 188, or polyethylene glycol 3350 based on polyoxyethylene (e.g., at concentrations of about 0.02% to about 0.1% (w / v), e.g., 0.03% (w / v)); and

[0655] Viscosity reducers, such as NaCl, MgCl2, D-arginine, L-arginine, or lysine (e.g., at a concentration of about 10-100 mM, such as 50 mM),

[0656] The pH of the formulation is approximately 5.0 to approximately 6.8 (e.g., 5.0-6.0 or 5.8).

[0657] In one embodiment of the present invention, the concentration of aflibercept in the aqueous pharmaceutical formulation is approximately 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. / 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; 137mg / ml; 134mg / ml; 135mg / ml; 136 ... g / ml; 138mg / ml; 139mg / ml; 140mg / ml; 141mg / ml; 142mg / ml; 143mg / ml; 144mg / ml; 145mg / ml; 146mg / ml; mg / ml; 152mg / ml; 153mg / ml; 154mg / ml; 155mg / ml; 156mg / ml; 157mg / ml; 158mg / ml; 159mg / ml; 160mg / ml; 5mg / ml; 166mg / ml; 167mg / ml; 168mg / ml; 169mg / ml; 170mg / ml; 171mg / ml; 172mg / ml; 173mg / ml; 174mg / ml; 79mg / 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; 203 mg / ml; 204mg / ml; 205mg / ml; 206mg / ml; 207mg / ml; 208mg / ml; 209mg / ml; 210mg / ml; 211mg / ml; 212mg / ml; 213mg / m l; 214mg / ml; 215mg / ml; 216mg / ml; 217mg / ml; 218mg / ml; 219mg / ml; 220mg / ml; 221mg / ml; 222mg / ml; 223mg / ml; 2 24mg / ml; 225mg / 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; 251mg / ml; 252mg / ml; 253mg / ml; 254mg / ml; 255 mg / 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 / ml; 269mg / ml; 270mg / ml; 271mg / ml; 272mg / ml; 273mg / ml; 274mg / ml; or 275mg / ml. ;

[0658] In one embodiment of the 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, with a pH of about 5.0 to about 6.8; wherein the aflibercept has less than about 3.5% of high molecular weight material immediately after production and purification and / or less than or equal to about 6% of high molecular weight material after being stored at about 2-8°C for about 24 months.

[0659] In one embodiment of the invention, the concentration of sucrose, mannitol, sorbitol, or trehalose is about 2-10% (w / v); the concentration of L-arginine is about 10-100 mM; the concentration of polysorbate 20 or polysorbate 80 is about 0.02-0.1% (w / v); and the concentration of the histidine-based buffer is about 5-25 mM; the pH is about 5.0 to about 6.8.

[0660] Treatment and application

[0661] The present invention provides a method for treating angiogenic eye diseases (e.g., neovascular age-related macular degeneration (nAMD)) by sequentially administering an initial loading dose (e.g., 2 mg or more, 4 mg or more, or preferably 8 mg or more of a VEGF antagonist or inhibitor, such as a VEGF receptor fusion protein like aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks), followed by additional doses every 12-20 weeks, preferably 12-16 weeks, 12 weeks, 16 weeks, or 20 weeks. For example, the present invention provides a method for treating or preventing vascular eye diseases such as neovascular age-related macular degeneration (nAMD), which involves sequentially administering one or more doses (e.g., 3, 4, or 5 times) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) of about 8 mg or more every 2-4 weeks or 3-5 weeks, for example, 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 12 weeks (or about every 3 months, or about every quarter, or about every 84 days), or every 16 weeks (or about every 4 months, or about every 1 / 3 year, or about every 112 days), or every 20 weeks. Dosing regimens including a 12-week third dosing interval may be referred to herein as a 12-week dosing regimen, 8q12, or HDq12; dosing regimens including a 16-week third dosing interval may be referred to herein as a 16-week dosing regimen, 8q16, or HDq16; and dosing regimens including a 20-week third dosing interval may be referred to herein as a 20-week dosing regimen, 8q20, or HDq20. Dosing regimens including a third dosing interval between 12 and 20 weeks may be referred to herein as a 12-20 week dosing regimen, 8q12-20, or HDq12-20.

[0662] Additionally, the present invention includes a method for treating angiogenic eye diseases (e.g., neovascular age-related macular degeneration (nAMD)) by administering ≥8 mg of VEGF receptor fusion protein, preferably aflibercept, once or multiple times every 4 weeks, 8 weeks, 12-20 weeks, 12-16 weeks, 12 weeks, or 16 weeks; and administering the first 3, 4, or 5 doses every 4 weeks, followed by administration approximately every 8 weeks.

[0663] In one embodiment of the invention, after a loading dose of ≥8 mg per month, the subject begins to receive a maintenance dose of ≥8 mg every 12, 16, or 20 weeks, without intermediate doses. The subject rapidly / immediately enters the maintenance dose phase after the loading dose phase. In another embodiment of the invention, the subject continues to receive ≥8 mg doses for 12 to 20 weeks, 12 weeks, 16 weeks, or 20 weeks, without any intermediate doses.

[0664] In one embodiment of the invention, the subject does not receive a dosing regimen modification (DRM) or discontinue treatment for at least 1, 2, 3, 4 or 5 years.

[0665] The terms “initial dose,” “second dose,” and “third dose” refer to the temporal sequence of administration of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept). Therefore, the “initial dose” is the dose administered at the start of the treatment regimen (also known as the “baseline dose”); the “second dose” is the dose administered after the initial dose; and the “third dose” is the dose administered after the second dose. The initial, second, and third doses may contain the same amount of VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), but typically differ in their frequency of administration. However, in some embodiments, the amount of VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) contained in the initial, second, and / or third doses differs from each other during treatment (e.g., adjusted upwards or downwards as appropriate).

[0666] Therefore, the dosing regimen of the present invention can be expressed as follows:

[0667] A method for treating angiogenic eye disease (e.g., nAMD) in a subject of need, the method comprising administering (e.g., intravitreal administration) to the subject of need.

[0668] A single initial dose of approximately ≥8 mg (approximately 100 μl or less, approximately 75 μl or less, or approximately 70 μl or less, e.g., 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) VEGF antagonists (e.g., VEGF receptor fusion proteins such as aflibercept) in the following amounts: 1; 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) followed by...

[0669] One or more (e.g., 2, 3, or 4 times, preferably 2 times) of a second dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0670] One or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept);

[0671] Each second dose is administered 2 to 4 weeks (preferably 4 weeks) immediately following the previous dose; and

[0672] Each third dose is administered at least 12, 16, or 20 weeks after the immediately preceding dose (preferably 12-20, 12-16, 12, or 16 weeks).

[0673] This invention includes methods in which one or more additional, unpredictable doses are administered to a subject in addition to any predetermined initial, second, and / or third dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept). Such doses are typically administered at the discretion of the treating physician based on the specific needs of the subject. This invention also provides a method for treating angiogenic ophthalmopathy by administering approximately ≥8 mg (e.g., approximately 100 μl or less, approximately 75 μl or less, or approximately 70 μl or less, such as 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) to a subject in need on a PRN-based basis. VEGF antagonists (e.g., VEGF receptor fusion proteins such as aflibercept) in the following μl; 71μl; 72μl; 73μl; 74μl; 75μl; 76μl; 77μl; 78μl; 79μl; 80μl; 81μl; 82μl; 83μl; 84μl; 85μl; 86μl; 87μl; 88μl; 89μl; 90μl; 91μl; 92μl; 93μl; 94μl; 95μl; 96μl; 97μl; 98μl; 99μl; or 100μl.

[0674] On-demand (PRN) treatment regimens require physician visits to be kept at fixed intervals (e.g., every 2, 3, 4, 8, 12, 16, or 20 weeks), and the decision to administer VEGF receptor fusion protein injections is based on the anatomical findings at each visit. Capped PRN dosing regimens require subjects to be treated at a minimum frequency, such as at least every 2, 3, or 4 months.

[0675] Treatment and extension (T&E) protocols require adjustments to the intervals between physician visits based on the patient's clinical course—for example, if the subject shows no signs of active disease (e.g., the macula remains dry and there is no leakage), the next one or more intervals can be extended; if there is fluid accumulation, the next interval will be shortened. At each visit following T&E, an injection of VEGF receptor fusion protein will be administered; the current clinical status only affects the duration of the next injection interval.

[0676] This invention includes embodiments in which, at any point during an HDq12-20, HDq12, HDq16, or HDq20 treatment regimen, a patient can switch to a PRN, capped PRN, or T&E regimen. The PRN, capped PRN, and / or T&E can continue indefinitely or can be stopped at any point in time, and then the HDq12, HDq16, or HDq20 regimen can be restarted at any stage. A period of PRN, capped PRN, and / or T&E can occur before or after any HDq12-20, HDq12, HDq16, or HDq20 regimen.

[0677] The method included in this invention comprises administering a desired dose of HDq12-20, HDq12, HDq16, or HDq20, wherein each third dose is administered 12-20 weeks, 12 weeks, 16 weeks, or 20 weeks after the immediate preceding dose, wherein the treatment interval between two third doses is extended (e.g., from 12 weeks to 13, 14, 15, 16, or 20 weeks, or from 16 weeks to 17, 18, 19, or 20 weeks) until signs of disease activity or visual impairment worsen or relapse, and then administration continues at the last third interval used or the penultimate third interval used. In one embodiment of the invention, the subject receives an initial, second interval, then a 12 or 16-week third interval, and then, after approximately one year, the third interval is extended to approximately 20 weeks.

[0678] The method included in this invention comprises administering a desired dose of HDq12-20 or HDq12 or HDq16 or HDq20, wherein the treatment interval between any two third doses is reduced (e.g., from 20 weeks to 8, 12 or 16 weeks, from 16 weeks to 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 weeks, or from 12 weeks to 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 weeks), for example, an improvement in signs of disease activity or visual impairment (e.g., stabilization or improvement of BCVA and / or stabilization or reduction of CRT), and thus, optionally, the dose interval may be extended, for example, extended back to a larger interval length.

[0679] For example, in one embodiment of the invention, the dosing interval, for example, during a dosing phase of 12-20 weeks, 12 weeks, or 16 weeks or 20 weeks, may be shortened (e.g., shortened to 8 weeks) if: • for example, the BCVA (ETDRS or Snellen equivalent) loss is greater than 5 letters (e.g., relative to the BCVA observed at approximately 12 weeks after treatment initiation), for example due to persistent or worsening AMD; and / or

[0680] • An increase in CRT greater than 25 micrometers is observed (e.g., relative to CRT observed approximately 12 weeks after treatment initiation), and / or if new foveal neovascularization or foveal hemorrhage is observed.

[0681] In one embodiment of the invention, if a subject receiving the HDq12 regimen meets the criteria for reducing the dose interval, the dose interval is reduced to 8 weeks at week 16 or 20.

[0682] In one embodiment of the invention, if a subject receiving the HDq16 regimen meets the criteria for reducing the dose interval, the dose interval is reduced to 8 weeks at week 16 or 20; and if a subject receiving the HDq16 regimen meets the criteria for reducing the dose interval, the dose interval is reduced to 12 weeks at week 24.

[0683] In one embodiment of the invention, the interval is not reduced to any interval shorter than 8 weeks.

[0684] For example, in one embodiment of the invention, the dosing interval, for example, during a dosing phase of 12-20 weeks, 12 weeks, 16 weeks, or 20 weeks, may be extended (e.g., increased by 4 weeks, for example, from 12 to 16 weeks, or 16 to 20 weeks), if:

[0685] • BCVA loss < 5 letters (e.g., from week 12 onwards);

[0686] • For example, there is no liquid in the central partition of the OCT, and / or

[0687] • No new foveal hemorrhage or foveal neovascularization.

[0688] (For example, if all three criteria are met)

[0689] See Figure 3 or Figure 4 .

[0690] The present invention also provides a method for treating angiogenic eye diseases (e.g., nAMD) by administering the following:

[0691] ≥8 mg approximately every 4, 5, 6, 7, 8, 12, 16, or 20 weeks (e.g., approximately 100 μl or less, approximately 75 μl or less, or approximately 70 μl or less, e.g., 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). ;70μl;71μl;72μl;73μl;74μl;75μl;76μl;77μl;78μl;79μl;80μl;81μl;82μl;83μl;84μl;85μl; 86μl; 87μl; 88μl; 89μl; 90μl; 91μl; 92μl; 93μl; 94μl; 95μl; 96μl; 97μl; 98μl; 99μl;; or 100μl) dose;

[0692] or

[0693] (For example, about ≥8 mg, such as 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; 7 A single initial dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) of 5 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) followed by

[0694] One or more (e.g., 2, 3, or 4 times (preferably 2 or 4 times)) of a second dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0695] One or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept);

[0696] Each second dose is administered 2 to 4 weeks (preferably 4 weeks) immediately following the previous dose; and

[0697] Each third dose is administered approximately 4, 5, 6, 7, or 8 weeks (e.g., 8 weeks) after the preceding dose.

[0698] or

[0699] Approximately every 4 weeks (q4w) ≥8 mg (e.g., approximately 100 μl or less, approximately 75 μl or less, or approximately 70 μl or less, e.g., 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). VEGF antagonists (e.g., VEGF receptor fusion proteins such as aflibercept) in the following amounts: 1; 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);

[0700] or

[0701] Over a period of approximately 48 weeks, administer approximately ≥8 mg (e.g., approximately 100 μl or less, approximately 75 μl or less, or approximately 70 μl or less, e.g., 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) at fewer than approximately 8 doses (e.g., approximately 5 or 6 doses) for approximately 48 weeks. VEGF antagonists (e.g., VEGF receptor fusion proteins such as aflibercept) in the following amounts: 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.

[0702] "Monthly" dosing or "dosing after one month" means dosing after about 28 days, about 4 weeks, or about 28 (±5 days) and may cover up to 5 weeks (±5 days).

[0703] Dosing every “4 weeks” or after “4 weeks” means dosing after about 28 days (±5 days), about one month or about 28 days (±5 days), and may cover dosing up to every 5 weeks (±5 days).

[0704] Dosing every “2-4 weeks” or after “2-4 weeks” means administering the drug after approximately 2 weeks (±5 days), 3 weeks (±5 days), or 4 weeks (±5 days).

[0705] Dosing every “8 weeks” or after “8 weeks” means dosing after approximately 2 months (±5 days) or approximately 56 (±5 days). Dosing every “12 weeks” or after “12 weeks” means dosing after approximately 3 months, approximately 84 days (±5 days), approximately 90 days (±5 days), or approximately 84 (±5 days).

[0706] Dosing every “16 weeks” or after “16 weeks” means dosing after approximately 4 months or approximately 112 days (±5 days). Dosing every “12–20 weeks” or after “12–20 weeks” means dosing after approximately 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks (±5 days), preferably after approximately 12–16 weeks (±5 days), approximately 12 weeks (±5 days), approximately 16 weeks (±5 days), or approximately 20 weeks (±5 days).

[0707] Dosing every "12-20 weeks" means administering the medication approximately at 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks (±5 days), preferably approximately 12-16 weeks (±5 days), approximately 12 weeks (±5 days), or approximately 16 weeks (±5 days).

[0708] Or administer the medication approximately 20 weeks (±5 days) later.

[0709] Doses of ≥8 mg cover doses of approximately 8 mg or more, such as approximately 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg.

[0710] In embodiments of the invention, any dosing frequency specified herein may be expressed as a specific frequency “±5 days” (e.g., where “4 weeks” is stated, the invention also includes embodiments such as 4 weeks ± 5 days). The term ± 5 days includes ±1, ±2, ±3, ±4 and / or ±5 days.

[0711] "Sequential administration" means administering each dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) to the eye of a subject at different times, such as on different dates at predetermined time intervals (e.g., hours, days, weeks, or months). The present invention includes a method comprising sequentially administering a single initial dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) to the eye of a subject, followed by one or more administrations of a second dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by one or more administrations of a third dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept).

[0712] An effective or therapeutically effective dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) for the treatment or prevention of angiogenic eye diseases refers to an amount of VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) sufficient to reduce one or more signs and / or symptoms of the disease or condition in the treated subject, whether by inducing the regression or elimination of such signs and / or symptoms or by inhibiting the progression of such signs and / or symptoms. In one embodiment of the invention, an effective or therapeutically effective dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) is approximately ≥8 mg per month, three times a month, followed by once every 12-20 weeks. In one embodiment of the invention, the reduction of signs and / or symptoms is, for example, achieving an increase of ≥5, 10, or 15 letters in BCVA (relative to baseline) by 1 year (e.g., an improvement of ≥5 letters in nAMD subjects and / or an improvement of 8-14 letters in DME patients / subjects); achieving BCVA ≥69 letters; achieving no fluid at the foveal center; a reduction in central retinal thickness (CRT) of about 150 micrometers or more (e.g., less than 300 micrometers in nAMD subjects / subjects; and / or a reduction of at least about 200 micrometers in DR or RVO patients / subjects) or achieving normal CRT (e.g., about 300 micrometers or less); and / or achieving no leakage on fluorescein angiography.

[0713] In one embodiment of the invention, subjects receiving HDq12-20, HDq16, or HDq16 receive fewer injections per month or quarter, for example, from the start of treatment up to week 60, compared to subjects receiving the 2q8 regimen. For example, subjects receiving HDq12 receive approximately 7 injections over 60 weeks and / or subjects receiving the HDq16 regimen receive approximately 6 injections over 60 weeks.

[0714] "Angiogenic eye disease" refers to any eye disease caused by or related to blood vessel growth or proliferation or vascular leakage. Non-limiting examples of angiogenic eye diseases that can be treated or prevented using the methods of the present invention include:

[0715] Age-related macular degeneration (neovascular (nAMD)),

[0716] Macular edema (ME)

[0717] • Macular edema following retinal vein occlusion (ME-RVO)

[0718] • Retinal vein occlusion (RVO)

[0719] Central retinal vein occlusion (CRVO),

[0720] Branch retinal vein occlusion (BRVO),

[0721] Diabetic macular edema (DME)

[0722] • Choroidal neovascularization (CNV)

[0723] ·Iris neovascularization,

[0724] Neovascular glaucoma,

[0725] • Postoperative fibrosis after glaucoma surgery

[0726] Proliferative vitreoretinopathy (PVR)

[0727] Optic disc neovascularization,

[0728] • Corneal neovascularization

[0729] •Retinal neovascularization,

[0730] • Vitreous neovascularization,

[0731] • Vascular obstruction,

[0732] • Polypoid choroidal vascular lesions (PCV), optionally, in which the subjects also had nAMD, • Pterygium,

[0733] Vascular retinopathy

[0734] • Diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (e.g., characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of approximately 47 or 53) or proliferative diabetic retinopathy; e.g., in subjects without DME), and

[0735] • Diabetic retinopathy in subjects with diabetic macular edema (DME).

[0736] This invention provides a method for treating angiogenic eye disease (e.g., nAMD) in subjects of need, comprising sequentially 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, such as a VEGF receptor fusion protein like aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks, preferably every 4 weeks; preferably three initial loading doses), followed by additional doses every 12-20 weeks, preferably 12-16 weeks, 12 weeks, 16 weeks, or 20 weeks, wherein the subject achieves and / or maintains this status after treatment initiation, for example at weeks 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.

[0737] • During the treatment process, BCVA adds, for example, the letters 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. ＞ 5. ＞ 10. ＞ 15 or ＞ 20 letters, or the absence of their loss, for example, as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart (e.g., at weeks 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);

[0738] • No dosage regimen modifications were accepted, for example, where the dosing interval (e.g., the interval between the third dose) was reduced from the HDq12-20 or HDq12 or HDq16 or HDq20 treatment regimen once started, for example, for at least 48, 60 or 96 weeks.

[0739] • Receive 100% of all predetermined doses, for example, for at least 48, 60, or 96 weeks;

[0740] • The average BCVA increases over a 12-week period, for example, by week 36 or week 48;

[0741] • No intraretinal fluid (IRF) and / or no subretinal fluid and / or subretinal epithelial fluid (e.g., at weeks 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), for example, where the subject had nAMD and PCV;

[0742] • The retina has no fluid in the central region (total fluid, intraretinal fluid [IRF] and / or subretinal fluid [SRF] and / or subretinal epithelial fluid) (e.g., from the start of treatment to approximately weeks 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); for example, where the subject has nAMD and PCV;

[0743] • No subretinal pigment epithelial fluid (e.g., from the start of treatment to approximately weeks 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);

[0744] • Fluorescein angiography (FA) without fluid leakage (e.g., approximately 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);

[0745] • The size of choroidal neovascularization (CNV) is reduced, for example, by approximately 2.9, 3, 3.5, 3.65, or 4 mm. 2 (For example, from the start of treatment to weeks 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), where the baseline size is approximately 6 or 6.5 mm. 2 ;

[0746] • The total damaged CNV area decreased compared to baseline (e.g., from the start of treatment to weeks 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).

[0747] • Loss of IRF and / or SRF, for example in the central partition (e.g. from the start of treatment to weeks 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);

[0748] • Reduction in central retinal thickness (CST) (e.g., from the start of treatment to weeks 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); for example, a reduction of approximately 125, 150, 175, or 200 micrometers;

[0749] • Improvement in vision-related quality of life, such as an improvement of approximately 3 or 4 points, as measured by the National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25) (e.g., from the start of treatment to weeks 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), for example, where the baseline score is approximately 76 or 77;

[0750] • Lack of adverse events (AEs) and / or serious AEs (SAEs) occurring during the treatment period, such as adverse events occurring during the treatment period that occurred at any time within 30 days of any injection, such as increased blood pressure, increased intraocular pressure, visual impairment, vitreous floaters, vitreous detachment, iris neovascularization and / or vitreous hemorrhage (e.g. from the start of treatment to weeks 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);

[0751] • An ETDRS letter score of at least 69 (approximately 20 / 40 Snellen equivalents) (e.g., from the start of treatment to weeks 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); and / or

[0752] • Efficacy and / or safety in subjects with nAMD were similar to those of aflibercept administered intravitreally at approximately 2 mg every 4 weeks for the first 3 months, followed by approximately every 8 weeks or every 2 months, for example, where efficacy was measured as an increase in BCVA and / or a decrease in central retinal thickness, and where safety was measured as the incidence of adverse events (adverse events occurring at any time within 30 days of any injection during the treatment period), such as increased blood pressure, increased intraocular pressure, visual disturbances, vitreous floaters, vitreous detachment, iris neovascularization, and / or vitreous hemorrhage.

[0753] This invention provides a method for treating angiogenic eye disease (e.g., nAMD) in subjects of need, comprising sequentially 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, such as a VEGF receptor fusion protein like aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks, preferably every 4 weeks; preferably three initial loading doses), followed by additional doses every 12-20 weeks, preferably 12-16 weeks, 12 weeks, 16 weeks, or 20 weeks, wherein the subject achieves and / or maintains this status after treatment initiation, for example at weeks 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.

[0754] • No anti-drug antibodies (e.g., anti-aflibercept) were detected during treatment;

[0755] • From the start of treatment to weeks 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, best corrected visual acuity (BVCA) improves;

[0756] • For example, an increase in BCVA measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent (e.g., from the start of treatment to weeks 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) ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or ≥7 letters;

[0757] • Improvement in BCVA 4 weeks after treatment initiation is approximately 2 or 3 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 3 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0758] • The improvement in BCVA at 8 weeks after the start of treatment is approximately 5 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 4 or 5 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0759] • The improvement in BCVA at 12 weeks after treatment initiation was approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0760] • Improvement in BCVA at 16 weeks after treatment initiation was approximately 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0761] • Improvement in BCVA at 20 weeks after treatment initiation was approximately 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0762] • Improvement in BCVA at 24 weeks after treatment initiation was approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0763] • Improvement in BCVA at 28 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0764] • Improvement in BCVA at 32 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0765] • Improvement in BCVA at 36 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0766] • Improvement in BCVA at 40 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0767] • Improvement in BCVA at 44 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0768] • Improvement in BCVA at 48 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0769] • Improvement in BCVA at 52 weeks after treatment initiation was approximately 7 or 8 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0770] • Improvement in BCVA at 56 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0771] • Improvement in BCVA at 60 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0772] • Improvement in BCVA at approximately 36 to approximately 48 weeks after treatment initiation is approximately 6 or 7 letters, or up to 46 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters, or up to 40 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0773] • Improvement in BCVA at approximately 48 to 60 weeks after treatment initiation is approximately 6 or 7 letters, or up to 46 letters (ETDRS or Snellen equivalent) with the HDq12 regimen; or approximately 6 or 7 letters, or up to 40 letters (ETDRS or Snellen equivalent) with the HDq16 regimen.

[0774] • Improvement in BCVA at 48 or 60 weeks after treatment initiation is at least about 5, 10, or 15 letters (ETDRS or Snellen equivalent) when using the HDq12 or HDq16 regimen.

[0775] • BCVA patients who were 48, 60, or 96 weeks post-treatment initiation did not lose 5, 10, or 15 letters (ETDRS letters or Snellen equivalents) when using the HDq12 or HDq16 regimens.

[0776] • Improvement in BCVA was observed around weeks 8, 9, 10, 11, or 12 after the start of treatment and was maintained during the treatment regimen (e.g., within approximately ±1 or ±2 ETDRS letters or Snellen equivalents), for example, until at least week 48.

[0777] • By approximately week 48 or 60, the BCVA should have at least 69 letters (ETDRS letters or Snellen equivalent) when receiving the HDq12 or HDq16 protocol;

[0778] • The BCVA at 4 weeks after treatment initiation is approximately 63 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 63 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0779] • The BCVA at 8 weeks after treatment initiation is approximately 65 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 65 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0780] • The BCVA at 12 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0781] • The BCVA at 16 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0782] • The BCVA at 20 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0783] • The BCVA at 24 weeks after treatment initiation is approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0784] • The BCVA at 28 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0785] • The BCVA at 32 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0786] • The BCVA at 36 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0787] • The BCVA at 40 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0788] • The BCVA at 44 weeks after treatment initiation is approximately 68 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0789] • The BCVA at 48 weeks after treatment initiation is approximately 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0790] • The BCVA between weeks 36 and 48 after the start of treatment is approximately 66, 67, 68, 69, 70, 71, or 72 letters or up to 91 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen.

[0791] • The BCVA between week 36 and week 48 after treatment initiation is approximately 66, 67, 68, 69, or 70 letters or up to 94 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0792] • The BCVA between weeks 48 and 60 after the start of treatment is approximately 66, 67, 68, 69, 70, 71, or 72 letters or up to 91 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen.

[0793] • The BCVA between weeks 48 and 60 after the start of treatment is approximately 66, 67, 68, 69, or 70 letters or up to 94 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0794] • By approximately week 48, the BCVA improvement is approximately 7, 8, 9, 10, or 11 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA... < 54 letters;

[0795] • By approximately week 48, the BCVA improves by about 5, 6, or 7 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA is about 55-73 letters;

[0796] • By approximately week 48, the BCVA improves by about 1, 2, or 3 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA is about ≥74 letters;

[0797] • By approximately week 48, the BCVA improved by about 4, 5, or 6 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA was about 63 letters and the baseline CRT was <400 micrometers;

[0798] • By approximately week 48, the BCVA improved by approximately 5, 6, 7, 8, or 9 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA was approximately 52 or 53 letters and the baseline CRT was ≥400 micrometers;

[0799] • By approximately week 48, the BCVA improved by approximately 2, 3, 4, 5, or 6 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA was approximately 56, 57, or 58 letters and the subject had the classic lowest CNV;

[0800] • By approximately week 48, the BCVA improved by about 5 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA was about 62, 63, or 64 letters and the subject had only occult CNV;

[0801] • By approximately week 48, the BCVA improved by about 6, 7, 8, 9, or 10 letters (ETDRS or Snellen equivalent), for example, where the baseline BCVA was about 53 or 54 letters and the subject had a classic-dominant CNV;

[0802] • The BCVA at 52 weeks after treatment initiation is approximately 67 or 68 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0803] • The BCVA at 56 weeks after treatment initiation is approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0804] • The BCVA at 60 weeks after treatment initiation is approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 or 67 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0805] • The BCVA between approximately 48 and 60 weeks after the start of treatment is approximately 66 to approximately 72 letters (ETDRS or Snellen equivalent) when using the HDq12 regimen; or approximately 66 to approximately 70 letters (ETDRS or Snellen equivalent) when using the HDq16 regimen.

[0806] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 4 after treatment initiation was approximately 59 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 64 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0807] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 8 after treatment initiation was approximately 63 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 65 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0808] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 12 after treatment initiation was approximately 63 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 66 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0809] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 16 after treatment initiation was approximately 64 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA at approximately 67 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0810] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 20 after treatment initiation was approximately 64 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 68 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0811] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 24 after treatment initiation was approximately 64 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 67 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0812] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 28 after treatment initiation was approximately 65 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 68 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0813] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 32 after treatment initiation was approximately 64 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 71 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0814] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 36 after treatment initiation was approximately 64 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 71 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0815] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 40 after treatment initiation was approximately 65 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 69 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0816] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 44 after treatment initiation was approximately 65 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 68 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0817] • In subjects with PCV and nAMD, BCVA increased by approximately 3.0 letters (ETDRS) at approximately week 4 after treatment initiation with the HDq12 regimen; and / or by approximately 3.0 letters with the HDq16 regimen;

[0818] • In subjects with PCV and nAMD, BCVA at approximately week 8 after treatment initiation increased by approximately 6.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 5.0 letters when receiving the HDq16 regimen;

[0819] • In subjects with PCV and nAMD, BCVA at approximately week 12 after treatment initiation increased by approximately 7.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 6.0 letters when receiving the HDq16 regimen;

[0820] • In subjects with PCV and nAMD, BCVA at approximately week 16 after treatment initiation increased by approximately 7.0 letters (ETDRS) when receiving the HDq12 regimen; and / or increased by approximately 7.0 letters when receiving the HDq16 regimen;

[0821] • In subjects with PCV and nAMD, BCVA at approximately week 20 after treatment initiation increased by approximately 7.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 7.0 letters when receiving the HDq16 regimen;

[0822] • In subjects with PCV and nAMD, BCVA at approximately week 24 after treatment initiation increased by approximately 7.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 5.0 letters when receiving the HDq16 regimen;

[0823] • In subjects with PCV and nAMD, BCVA at approximately week 28 after treatment initiation increased by approximately 9.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 7.0 letters when receiving the HDq16 regimen;

[0824] • In subjects with PCV and nAMD, BCVA at approximately week 32 after treatment initiation increased by approximately 8.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 10.0 letters when receiving the HDq16 regimen;

[0825] • In subjects with PCV and nAMD, BCVA at approximately week 36 after treatment initiation increased by approximately 8.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 10.0 letters when receiving the HDq16 regimen;

[0826] • In subjects with PCV and nAMD, BCVA at approximately week 40 after treatment initiation increased by approximately 9.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 8.0 letters when receiving the HDq16 regimen;

[0827] • In subjects with PCV and nAMD, BCVA at approximately week 44 after treatment initiation increased by approximately 9.0 letters (ETDRS) when receiving the HDq12 regimen; and / or by approximately 8.0 letters when receiving the HDq16 regimen;

[0828] • In subjects with PCV and nAMD, BCVA at approximately week 48 after treatment initiation increased by approximately 9.0 letters (ETDRS) when receiving the HDq12 regimen; and / or increased by approximately 9.0 letters when receiving the HDq16 regimen;

[0829] • In subjects with polypoid choroidal vascular disease (PCV) and neovascular age-related macular degeneration (nAMD), the BCVA at approximately week 48 after treatment initiation was approximately 65 letters (ETDRS) when receiving the HDq12 regimen (e.g., where the baseline BCVA was approximately 57 letters); and / or the BCVA was approximately 69 letters (ETDRS) when receiving the HDq16 regimen (e.g., where the baseline BCVA was approximately 60 letters).

[0830] • The reduction in total lesion size at approximately week 12 after treatment initiation was approximately 0.45 (±2.3) or 0.5 (±2.3) or 0.23 mm when receiving the HDq12 regimen. 2 or at most 11mm 2 Or, when accepting the HDq16 scheme, approximately 0.40 (±2.6) or 0.13 mm. 2 or at most 17mm 2 For example, the baseline total lesion area is approximately 5, 6, or 7 mm. 2 ;

[0831] • The reduction in total lesion size at approximately week 48 after treatment initiation was approximately 0.36, 0.4, 0.46, or 0.26 (±2.9 or ±0.19) mm when receiving the HDq12 regimen. 2 Or, when accepting the HDq16 scheme, approximately 0.2, 0.2856, 0.3, or 0.04 (±3.2 or ±0.2) mm. 2 or at most 16mm 2 For example, the baseline total lesion area is approximately 5, 6, or 7 mm. 2 ;

[0832] • The reduction in total lesion size at approximately week 60 after treatment initiation was approximately 0.52, 0.6, or 0.32 (±2.8) mm when receiving the HDq12 regimen. 2 or at most 14mm 2 Or, when accepting the HDq16 scheme, approximately 0.35, 0.4, or 0.14 (±3.2) mm. 2 or at most 17mm 2 For example, the baseline total lesion area is approximately 5, 6, or 7 mm. 2 ;

[0833] • Reduction in central retinal thickness (CRT) of at least approximately 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 micrometers (e.g., from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48);

[0834] • The change in central retinal thickness 4 weeks after the start of treatment is approximately -120, -121, -122, -122.4, or -120.2 micrometers when using the HDq12 protocol; or approximately -126, -127, -126.6, or -126.3 micrometers when using the HDq16 protocol;

[0835] • The change in central retinal thickness 8 weeks after the start of treatment is approximately -132, -133, -134, -135, -136, -136.2, or -132.8 micrometers when using the HDq12 protocol; or approximately -139, -140, -139.5, or -139.6 micrometers when using the HDq16 protocol;

[0836] • The change in central retinal thickness 12 weeks after the start of treatment was approximately -136, -137, -138, -139, -140, -141, -140.9, or -136.6 micrometers when using the HDq12 protocol; or approximately -144, -143, or -143.5 micrometers when using the HDq16 protocol;

[0837] • The change in central retinal thickness 16 weeks after treatment initiation was approximately -120, -121, -122, -123, -124, -123.4, or -120.1 micrometers when using the HDq12 protocol; or approximately -132, -133, -132.1, or -133.1 micrometers when using the HDq16 protocol;

[0838] • The change in central retinal thickness 20 weeks after the start of treatment is approximately -110, -111, -112, -113, -114, -113.6, or -110.9 micrometers when using the HDq12 protocol; or approximately -115, -116, -117, -118, -115.8, or -117.7 micrometers when using the HDq16 protocol;

[0839] • The change in central retinal thickness 24 weeks after the start of treatment was approximately -134, -135, -136, -137, -138, -137.6, or -134.9 micrometers when using the HDq12 protocol; or approximately -105, -106, -107, -108, -105.3, or -107.8 micrometers when using the HDq16 protocol;

[0840] • The change in central retinal thickness 28 weeks after the start of treatment was approximately -130, -131, -132, -133, -134, -133.7, or -130.7 micrometers when using the HDq12 protocol; or approximately -144, -145, -146, -147, -148, -144.7, or -147.2 micrometers when using the HDq16 protocol;

[0841] • The change in central retinal thickness 32 weeks after the start of treatment was approximately -118 or -19 or -120 or -121 or -120.4 or -118.1 micrometers when using the HDq12 protocol; or approximately -141 or -142 or -143 or -144 or -141.5 or -144 micrometers when using the HDq16 protocol;

[0842] • The change in central retinal thickness 36 weeks after the start of treatment was approximately -142, -143, -144, -144.2, or -142.2 micrometers when using the HDq12 protocol; or approximately -126, -127, -128, -129, -130, -131, -126.4, or -130.5 micrometers when using the HDq16 protocol;

[0843] • The change in central retinal thickness 40 weeks after the start of treatment is approximately -131, -132, -133, -134, -133.8, or -131.2 micrometers when using the HDq12 protocol; or approximately -127, -128, or -127.5 micrometers when using the HDq16 protocol;

[0844] • The change in central retinal thickness 44 weeks after the start of treatment was approximately -120, -121, -122, -123, -124, -125, -124.7, or -120.3 micrometers when using the HDq12 protocol; or approximately -143, -144, -145, or -144.8 micrometers when using the HDq16 protocol;

[0845] • Changes in central retinal thickness at 48 weeks after treatment initiation were approximately -141, -141.9, -142, -142.3, -143, -144, -144.4, -145, -146, -147, -147.37, or -148 micrometers with the HDq12 regimen (e.g., where the baseline CRT is approximately 370 micrometers); or approximately -143, -144, -145, -146, -146.76, -147, -147.1, -148, -143.8, or -147.1 micrometers with the HDq16 regimen (e.g., where the baseline CRT is approximately 370 micrometers).

[0846] • The change in central retinal thickness 60 weeks after the start of treatment was approximately -153 micrometers when using the HDq16 regimen;

[0847] • The central retinal thickness at week 4 after treatment initiation was approximately 248.2 micrometers with the HDq12 protocol; or approximately 244.1 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0848] • The central retinal thickness at week 8 after treatment initiation was approximately 234.4 micrometers with the HDq12 protocol; or approximately 231.2 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0849] • The central retinal thickness at week 12 after treatment initiation was approximately 229.7 micrometers with the HDq12 protocol; or approximately 226.7 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0850] • The central retinal thickness at week 16 after treatment initiation was approximately 247.2 micrometers with the HDq12 protocol; or approximately 238.6 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0851] • The central retinal thickness at week 20 after treatment initiation was approximately 257 micrometers with the HDq12 protocol; or approximately 254.9 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers);

[0852] • The central retinal thickness at week 24 after treatment initiation was approximately 233 micrometers with the HDq12 protocol; or approximately 265.4 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers);

[0853] • The central retinal thickness at week 28 after treatment initiation was approximately 236.9 micrometers with the HDq12 protocol; or approximately 226 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0854] • At week 32 after treatment initiation, the central retinal thickness was approximately 250.2 micrometers with the HDq12 protocol; or approximately 229.2 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0855] • At week 36 after treatment initiation, the central retinal thickness was approximately 226.4 micrometers with the HDq12 protocol; or approximately 244.3 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0856] • The central retinal thickness at week 40 after treatment initiation was approximately 236.8 micrometers with the HDq12 protocol; or approximately 243.7 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0857] • The central retinal thickness at week 44 after treatment initiation was approximately 245.9 micrometers with the HDq12 protocol; or approximately 227.7 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0858] • The central retinal thickness at week 48 after treatment initiation was approximately 226 micrometers with the HDq12 protocol; or approximately 226.9 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers);

[0859] • At week 52 after treatment initiation, the central retinal thickness was approximately 227.4 micrometers with the HDq12 protocol; or approximately 231.1 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0860] • The central retinal thickness at week 56 after treatment initiation was approximately 234.3 micrometers with the HDq12 protocol; or approximately 233.2 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0861] • The central retinal thickness at week 60 after treatment initiation was approximately 218.8 micrometers with the HDq12 protocol; or approximately 221.9 micrometers with the HDq16 protocol (e.g., where the initial CRT was approximately 370 micrometers).

[0862] • When using the HDq12 regimen, the central retinal thickness changed by approximately -112, -113, or -112.4 micrometers from week 0 (treatment start) to week 4;

[0863] • When using the HDq12 protocol, the central retinal thickness changed by approximately -13, -14, or -13.8 micrometers between weeks 4 and 8;

[0864] • When using the HDq12 protocol, the central retinal thickness variation between week 8 and week 12 was approximately -4, -5, or -4.7 micrometers;

[0865] • When using the HDq12 protocol, the central retinal thickness change was approximately -24 or -25 micrometers between weeks 20 and 24;

[0866] • When using the HDq12 protocol, the central retinal thickness variation between week 32 and week 36 was approximately -23, -24, or -23.8 micrometers;

[0867] • When using the HDq12 protocol, the central retinal thickness variation between week 44 and week 48 was approximately -19, -20, or -19.7 micrometers;

[0868] • When using the HDq16 protocol, the central retinal thickness varies by approximately -126, -127, or -126.6 micrometers between week 0 and week 4;

[0869] • When using the HDq16 protocol, the central retinal thickness varied by approximately -12, -13, or -12.9 micrometers between weeks 4 and 8;

[0870] • When using the HDq16 protocol, the central retinal thickness variation was approximately -4, -5, or -4.5 micrometers between weeks 8 and 12;

[0871] • When using the HDq16 protocol, the central retinal thickness varied by approximately -39, -40, or -39.4 micrometers between weeks 24 and 28;

[0872] • When using the HDq16 protocol, the central retinal thickness variation was approximately 0, -1, or -0.6 micrometers between weeks 36 and 40;

[0873] • When using the HDq16 protocol, the central retinal thickness change was approximately -15 or -16 micrometers between weeks 40 and 44;

[0874] • When using the HDq16 protocol, the central retinal thickness variation was approximately 0, -1, or -0.8 micrometers between weeks 44 and 48;

[0875] • When using the HDq16 protocol, the central retinal thickness varied by approximately -11, -12, or -11.3 micrometers between weeks 56 and 60;

[0876] • The CRT decreases at approximately 4, 5, 6, 7 or 8 weeks after treatment initiation, and is maintained thereafter during the treatment regimen (e.g., within approximately ±10, ±11 or ±12 micrometers), for example, until at least week 48.

[0877] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately week 4 after treatment initiation was approximately -130.2 micrometers when receiving the HDq12 regimen; and / or approximately -111.6 micrometers when receiving the HDq16 regimen;

[0878] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately week 8 after treatment initiation was approximately -152.5 micrometers when receiving the HDq12 regimen; and / or approximately -134.5 micrometers when receiving the HDq16 regimen;

[0879] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 12 weeks after treatment initiation was approximately -160.9 micrometers when receiving the HDq12 regimen; and / or approximately -147.7 micrometers when receiving the HDq16 regimen;

[0880] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 16 weeks after treatment initiation was approximately -150 micrometers when receiving the HDq12 regimen; and / or approximately -141.1 micrometers when receiving the HDq16 regimen;

[0881] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 20 weeks after treatment initiation was approximately -135.9 micrometers when receiving the HDq12 regimen; and / or approximately -128 micrometers when receiving the HDq16 regimen;

[0882] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 24 weeks after treatment initiation was approximately -162.7 micrometers when receiving the HDq12 regimen; and / or approximately -115.7 micrometers when receiving the HDq16 regimen;

[0883] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 28 weeks after treatment initiation was approximately -163.4 micrometers when receiving the HDq12 regimen; and / or approximately -147.9 micrometers when receiving the HDq16 regimen;

[0884] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 32 weeks after treatment initiation was approximately -148.2 micrometers when receiving the HDq12 regimen; and / or approximately -142.3 micrometers when receiving the HDq16 regimen;

[0885] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 36 weeks after treatment initiation was approximately -167.4 micrometers when receiving the HDq12 regimen; and / or approximately -132.6 micrometers when receiving the HDq16 regimen;

[0886] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 40 weeks after treatment initiation was approximately -159.2 micrometers when receiving the HDq12 regimen; and / or approximately -121.1 micrometers when receiving the HDq16 regimen;

[0887] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 44 weeks after treatment initiation was approximately -146.7 μm when receiving the HDq12 regimen; and / or approximately -143.9 μm when receiving the HDq16 regimen;

[0888] • In subjects with PCV and nAMD, the change in central retinal thickness (CRT) at approximately 48 weeks after treatment initiation was approximately -170.6 micrometers when receiving the HDq12 regimen; and / or approximately -146.1 micrometers when receiving the HDq16 regimen;

[0889] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately week 4 after treatment initiation was approximately 260.2 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 263.6 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0890] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately week 8 after treatment initiation was approximately 237.9 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 240.7 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0891] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 12 weeks after treatment initiation was approximately 229.5 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 227.5 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0892] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 16 weeks after treatment initiation was approximately 240.4 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 234.1 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0893] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 20 weeks after treatment initiation was approximately 254.5 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 247.2 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0894] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 24 weeks after treatment initiation was approximately 227.7 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 259.5 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0895] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 28 weeks after treatment initiation was approximately 227 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 227.3 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0896] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 32 weeks after treatment initiation was approximately 242.2 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 232.9 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0897] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 36 weeks after treatment initiation was approximately 223 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 242.6 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0898] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 40 weeks after treatment initiation was approximately 231.2 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 254.1 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0899] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 44 weeks after treatment initiation was approximately 243.7 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 231.3 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0900] • In subjects with PCV and nAMD, the central retinal thickness (CRT) at approximately 48 weeks after treatment initiation was approximately 219.8 micrometers when receiving the HDq12 regimen (e.g., where the baseline CRT was approximately 390.4 micrometers); and / or approximately 229.1 micrometers when receiving the HDq16 regimen (e.g., when the baseline CRT was approximately 375.2 micrometers);

[0901] • The reduction in choroidal neovascularization size at week 12 was approximately 0.6, 1.5, 1, or 2 mm when receiving the HDq12 regimen. 2 (±3.7mm 2 ), or at most about 11 or 12 mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 5, or 6 mm) 2 hour);

[0902] • The reduction in choroidal neovascularization size at week 48 was approximately 2, 3, 4, 3.5, or 2.5 mm when receiving the HDq12 regimen. 2 (±5mm 2 ), or at most about 15 or 16 mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 5, or 6 mm) 2 hour);

[0903] • The reduction in choroidal neovascularization size at week 60 was approximately 2, 3, 4, 3.8, and 2.8 mm when receiving the HDq12 regimen. 2 (±5mm 2 ), or at most 13mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 5, or 6 mm) 2 hour);

[0904] • The reduction in choroidal neovascularization size at week 12 was approximately 1, 2, 0.5, or 1.5 mm when receiving the HDq16 regimen. 2 (±4.2mm 2 ), or at most 17mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 4.5, 5, 6, 6.5, or 7 mm) 2 hour);

[0905] • Choroidal neovascularization size at week 48 (mm) 2 The reduction in size is approximately 1, 2, 1.4, or 3 mm when the HDq16 scheme is accepted. 2 (±5mm 2 ), or at most 16mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 4.5, 5, 6, 6.5, or 7 mm) 2 hour);

[0906] • The reduction in choroidal neovascularization size at week 60 was approximately 2, 3, 4, or 3.7 mm when receiving the HDq16 regimen. 2 (±5.7mm 2 ), or at most 17mm 2 (For example, when the baseline size at the start of treatment is approximately 4, 4.5, 5, 6, 6.5, or 7 mm) 2 hour);

[0907] • At baseline, the concentration of free aflibercept in plasma is approximately 0 mg / L (or <0.0156 mg / L);

[0908] • Approximately 4 hours after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.04 (±0.06) or 0 mg / L (or <0.0156 mg / L);

[0909] • Approximately 8 hours after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.05 (±3.8), 0.1 (±0.102), or 0.067 mg / L, respectively.

[0910] • On approximately day 2 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.11 (±2.21), 0.146 (±0.110), or 0.0903 mg / L;

[0911] • Around day 3 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.11 (±2.06), 0.137 (±0.0947), or 0.112 mg / L;

[0912] • Around day 5 after the start of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.08 (±1.86), 0.09 (±0.048), or 0.085 mg / L, respectively.

[0913] • Around day 8 after the start of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.07 (±1.75), 0.0794 (±0.041), or 0.068 mg / L;

[0914] • Around day 15 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.04 (±1.76), 0.044 (±0.0199), or 0.039 mg / L;

[0915] • Around day 22 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.02 (±1.76), 0.021 (±0.0148), or 0.023 mg / L;

[0916] • Around day 29 after the initiation of HDq12 or HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.0077 (±0.0096) or 0 mg / L (or <0.0156 mg / L);

[0917] • Approximately 48 weeks after the initiation of HDq12 treatment, the plasma concentration of free aflibercept was approximately 0.014 mg / L (±0.018).

[0918] • Approximately 48 weeks after the initiation of HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.00056 mg / L (±0.0038).

[0919] • Approximately 28 weeks after the initiation of HDq12 treatment, the plasma concentration of free aflibercept was approximately 0.00088 mg / L (±0.0053).

[0920] • Approximately 28 weeks after the initiation of HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.015 mg / L (±0.016);

[0921] • Approximately 12 weeks after the initiation of HDq12 treatment, the plasma concentration of free aflibercept was approximately 0.02 mg / L (±2) or approximately 0.028 mg / L (±0.023);

[0922] • Approximately 12 weeks after the initiation of HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.02 mg / L (±2) or approximately 0.029 mg / L (±0.023);

[0923] • Approximately 4 weeks after the initiation of HDq12 treatment, the plasma concentration of free aflibercept was approximately 0.017 mg / L (±0.016);

[0924] • Approximately 4 weeks after the initiation of HDq16 treatment, the plasma concentration of free aflibercept was approximately 0.017 mg / L (±0.016);

[0925] • Approximately 4 hours after administration of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept is approximately 0.00 mg / L;

[0926] • Approximately 8 hours after administration of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept is approximately 0.00 mg / L;

[0927] • On approximately day 2 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.06 (±3.50) or 0.12 (±0.19) mg / L;

[0928] • On approximately day 3 following the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.13 (±2.07) or 0.17 (±0.155) mg / L.

[0929] • On approximately day 5 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.18 (±1.9) or 0.22 (±0.157) mg / L;

[0930] • Around day 8 after the initiation of HDq12 or HDq16 treatment, the adjusted plasma concentration of bound aflibercept was approximately 0.31 (±1.56) or 0.33 (±0.135) mg / L;

[0931] • On d...

Claims

1. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering one or more doses of aflibercept at certain intervals and amounts, whereby After intravitreal injection of aflibercept, the clearance of free aflibercept from the ocular chamber was approximately 0.3 mL / day, 0.4 mL / day, 0.41 mL / day, or 0.37-0.46 mL / day. The amount of free aflibercept in the ocular chamber of the subject reaches the lower limit of quantitation (LLOQ) at about 15 weeks following the intravitreal injection of aflibercept; and / or The time to reach the lower limit of quantification (LLOQ) of free aflibercept in the subject's plasma following the intravitreal injection of aflibercept is about 3.8 weeks, 3.5 weeks, or 3.5-3.8 weeks.

2. A method for slowing the clearance of free aflibercept from the ocular chamber after intravitreal injection relative to the clearance rate of aflibercept from the ocular chamber after intravitreal injection of 2 mg or <4 mg aflibercept, the method comprising Intravitreal injection into the eye of a subject in need A single initial dose of aflibercept of about 8 mg or more, followed by One or more second doses of aflibercept of about 8 mg or more, followed by one or more third doses of aflibercept of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

3. The method of any one of claims 1 to 2, wherein the clearance of free aflibercept from the ocular chamber is about 34% slower than the clearance of free aflibercept from the ocular chamber following intravitreal injection of 2 mg or <4 mg aflibercept.

4. The method of any one of claims 1 to 3, wherein the clearance of free aflibercept from the ocular chamber following intravitreal injection of >8 mg aflibercept is about 0.37-0.46 mL / day or 0.41 mL / day.

5. A method for increasing the duration of efficacy of free aflibercept and / or the time at which the amount of free aflibercept in the ocular chamber of a subject reaches the lower limit of quantitation (LLOQ) after intravitreal injection of aflibercept relative to the time at which the amount of free aflibercept in the ocular chamber of a subject reaches the LLOQ after intravitreal injection of about 2 mg or <4 mg of aflibercept, the method comprising intravitreally injecting into an eye of a subject in need thereof A single initial dose of aflibercept of about 8 mg or more, followed by One or more second doses of aflibercept of about 8 mg or more, followed by one or more third doses of aflibercept of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

6. The method of claim 5, wherein the duration of efficacy of free aflibercept and / or the time until the amount of free aflibercept in the ocular chamber of the subject reaches the lower limit of quantitation (LLOQ) after the intravitreal injection of aflibercept is increased by about 5 weeks or 6 weeks relative to the time until the amount of free aflibercept in the ocular chamber of the subject reaches the LLOQ after the intravitreal injection of about 2 mg or <4 mg of aflibercept.

7. The method of any one of claims 5 to 6, wherein the time for the amount of free aflibercept in the ocular chamber of the subject to reach the lower limit of quantitation (LLOQ) after the intravitreal injection of aflibercept is increased by greater than about 1 week, 2 weeks, 1.2 weeks or 1.3 weeks relative to the time for free aflibercept in the ocular chamber of the subject to reach LLOQ after intravitreal injection of about 2 mg or <4 mg aflibercept.

8. The method of any one of claims 5 to 7, wherein the time to reach the lower limit of quantitation (LLOQ) of the amount of free aflibercept in the ocular chamber of the subject following the intravitreal injection of >8 mg aflibercept is about 15 weeks.

9. The method of any one of claims 5 to 8, wherein the time to reach the lower limit of quantitation (LLOQ) of the amount of free aflibercept in the ocular chamber of the subject following the intravitreal injection of >8 mg aflibercept is greater than about 8 weeks, 8.7 weeks, 8.71 weeks, 9 weeks, 9.2 weeks, 9.21 weeks, or 10 weeks.

10. A method for increasing the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the plasma of a subject following intravitreal injection of aflibercept relative to the time to reach the LLOQ of free aflibercept in the plasma of a subject following intravitreal injection of about 2 mg or <4 mg of aflibercept, the method comprising intravitreally injecting into an eye of a subject in need thereof A single initial dose of aflibercept of about 8 mg or more, followed by One or more second doses of aflibercept of about 8 mg or more, followed by one or more third doses of aflibercept of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

11. The method of claim 10, wherein the LLOQ of free aflibercept measured in plasma is about 0.0156 mg / L.

12. The method of any one of claims 10 to 11, wherein the time for free aflibercept to reach the lower limit of quantitation (LLOQ) in the subject's plasma following the intravitreal injection of aflibercept is increased by about 2 weeks relative to the time for free aflibercept to reach the LLOQ in the subject's plasma following an intravitreal injection of about 2 mg of aflibercept.

13. The method of any one of claims 10 to 12, wherein the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the subject's plasma following the intravitreal injection of >8 mg aflibercept is about 3 weeks, 3.5 weeks, 3.8 weeks or 4 weeks.

14. The method of any one of claims 10 to 13, wherein the time to reach the lower limit of quantitation (LLOQ) of free aflibercept in the subject's plasma following the intravitreal injection of >8 mg aflibercept is greater than about 1.5 weeks or 1.6 weeks.

15. The method of any one of claims 1 to 14, wherein <4 mg is about 2 mg or 2-4 mg.

16. The method of any one of claims 1 to 15, wherein the subject suffers from neovascular age-related macular degeneration (nAMD).

17. The method of any one of claims 1 to 16, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising a histidine-based buffer.

18. The method of any one of claims 1 to 17, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising arginine.

19. The method of any one of claims 1 to 18, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation at a pH of about 5.

8.

20. The method of any one of claims 1 to 19, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising a sugar or a polyol.

21. The method of any one of claims 1 to 20, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising sucrose.

22. The method of any one of claims 1 to 21, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation, wherein the aflibercept has less than about 3.5% high molecular weight species immediately after production and purification and / or has less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months.

23. The method of any one of claims 1 to 22, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation, the aqueous pharmaceutical formulation comprising an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein, the fusion protein comprising two polypeptides, each polypeptide comprising an immunoglobulin-like (Ig) domain 2 of VEGFR1, an Ig domain 3 of VEGFR2, and a multimerization component; about 10 mM-100 mM L-arginine; sucrose; a histidine-based buffer; and a surfactant; wherein the pH of the formulation is about 5.0 to about 6.8; wherein the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification and / or has less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months.

24. A method for - for the treatment or prevention of neovascular age-related macular degeneration (nAMD) in a subject in need thereof, - for improving best corrected visual acuity in subjects in need with nAMD; or - For promoting retinal dryness in subjects in need thereof with nAMD; The method comprises administering one or more doses of about 8 mg or more of a VEGF receptor fusion protein to the eye of the subject once every 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks, or 12-20 weeks, or 12-16 weeks, or 16-20 weeks.

25. The method of any one of claims 1 to 24 for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

26. The method of any one of claims 1 to 24 for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12 weeks after the immediately preceding dose.

27. The method of any one of claims 1 to 24 for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye a single initial dose of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 16 weeks after the immediately preceding dose.

28. The method of any one of claims 1 to 24 for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 20 weeks after the immediately preceding dose.

29. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering 8 mg VEGF receptor fusion protein (0.07 mL or 70 microliters) via intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by 8 mg aflibercept (0.07 mL) via intravitreal injection once every 8 to 16 weeks (2-4 months + / - 7 days).

30. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering 8 mg VEGF receptor fusion protein (0.07 mL or 70 microliters) via intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by administration of 8 mg VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 12 weeks (2-4 months + / - 7 days).

31. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering 8 mg VEGF receptor fusion protein (0.07 mL or 70 microliters) via intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by administration of 8 mg VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 16 weeks (2-4 months + / - 7 days).

32. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering 8 mg VEGF receptor fusion protein (0.07 mL or 70 microliters) via intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by administration of 8 mg VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 20 weeks (+ / - 7 days).

33. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof with a dose of 8 mg of aflibercept (equivalent to 70 microliters of injection), the method comprising administering 1 injection per month (every 4 weeks) for 3 consecutive doses of said 8 mg aflibercept, wherein the injection interval can then be extended to every 16 weeks based on the physician's judgment of visual and / or anatomical results.

34. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, in: (1) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein, and then the method includes administering the initial dose of 8 mg of VEGF receptor fusion protein to the subject one month later, and administering the first second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and administering the second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen; (2) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein, and then the method comprises administering to the subject a first second dose of 8 mg of VEGF receptor fusion protein one month later, and administering a second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (3) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, and the method then comprises administering to the subject a second dose of 8 mg of VEGF receptor fusion protein one month later, and then administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen; (4) the subject has received an initial 2 mg dose of VEGF receptor fusion protein, and then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein one month later, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (5) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first second dose of 2 mg of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject an initial dose of 8 mg of VEGF receptor fusion protein one month later, and administering a first second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and administering a second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen; (6) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first second dose of 2 mg of VEGF receptor fusion protein one month later, and then the method includes administering to the subject a first second dose of 8 mg of VEGF receptor fusion protein one month later, and administering a second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (7) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein one month later, and then, every 12 weeks, 16 weeks, or 20 weeks thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing regimen; (8) The subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein 1 month later, and then the method includes administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein 1 month later, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (9) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later and received a second 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject an initial dose of 8 mg of VEGF receptor fusion protein one month later, and administering a first 8 mg second dose of VEGF receptor fusion protein one month thereafter; and administering a second 8 mg second dose of VEGF receptor fusion protein one month thereafter; then, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks thereafter according to the HDq12, HDq16 or HDq20 dosing regimen; (10) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first second dose of 2 mg of VEGF receptor fusion protein one month later and a second second dose of 2 mg of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a first second dose of 8 mg of VEGF receptor fusion protein one month later and a second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks according to the HDq12, HDq16 or HDq20 dosing regimen; (11) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first second dose of 2 mg of VEGF receptor fusion protein one month later and received a second second dose of 2 mg of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a second second dose of 8 mg of VEGF receptor fusion protein one month later, and then, every 12 weeks, 16 weeks or 20 weeks thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein according to the HDq12, HDq16 or HDq20 dosing regimen; (12) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein 1 month later and received a second 2 mg second dose of VEGF receptor fusion protein 1 month later, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein 2 months later, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (13) The subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received the first 2 mg second dose of VEGF receptor fusion protein after 1 month and received the second 2 mg second dose of VEGF receptor fusion protein after 1 month and received the first 2 mg maintenance dose of VEGF receptor fusion protein after 8 weeks, then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein up to 2 months after the last dose of VEGF receptor fusion protein, and administering the first 8 mg second dose of VEGF receptor fusion protein 1 month thereafter; and administering the second 8 mg second dose of VEGF receptor fusion protein 1 month thereafter; then, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks thereafter according to the HDq12, HDq16 or HDq20 dosing regimen; (14) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later and received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later, and the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein up to two months after the last dose of VEGF receptor fusion protein, and administering a second 8 mg second dose of VEGF receptor fusion protein one month thereafter, and then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks according to the HDq12, HDq16 or HDq20 dosing regimen; (15) the subject has received an initial dose of 2 mg of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein one month later and received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a first 2 mg maintenance dose of VEGF receptor fusion protein eight weeks later, and the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein up to two months after the last dose of VEGF receptor fusion protein, and then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks according to the HDq12, HDq16 or HDq20 dosing regimen; (16) the subject has received an initial 2 mg dose of VEGF receptor fusion protein and received a first 2 mg second dose of VEGF receptor fusion protein 1 month later and received a second 2 mg second dose of VEGF receptor fusion protein 1 month later and received a first 2 mg maintenance dose of VEGF receptor fusion protein 8 weeks later, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein up to 2 months after the last dose of VEGF receptor fusion protein, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; in, (i) The HDq12 dosing regimen includes: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 weeks after the immediately preceding dose; (ii) The HDq16 dosing regimen comprises: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered approximately 16 weeks after the immediately preceding dose; and (iii) The HDq20 dosing regimen comprises: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 20 weeks after the immediately preceding dose.

35. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, in: (a) the subject has received an initial dose of 8 mg of VEGF receptor fusion protein, and then the method comprises administering to the subject a first second dose of 8 mg of VEGF receptor fusion protein one month later and a second second dose of 8 mg of VEGF receptor fusion protein one month thereafter; and then, thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (b) the subject has received an initial dose of 8 mg of VEGF receptor fusion protein and received a first second dose of 8 mg of VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a second second dose of 8 mg of VEGF receptor fusion protein one month later, and then, every 12 weeks, 16 weeks or 20 weeks thereafter, administering one or more maintenance doses of 8 mg of VEGF receptor fusion protein according to the HDq12, HDq16 or HDq20 dosing regimen; (c) the subject has received an initial 8 mg dose of VEGF receptor fusion protein and received a first 8 mg second dose of VEGF receptor fusion protein one month later and received a second 8 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein after 12 weeks, 16 weeks or 20 weeks, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks or 20 weeks according to the HDq12, HDq16 or HDq20 dosing regimen; or (d) the subject has received an initial 8 mg dose of VEGF receptor fusion protein and received a first 8 mg second dose of VEGF receptor fusion protein one month later and received a second 8 mg second dose of VEGF receptor fusion protein one month later, and then every 12 weeks, 16 weeks, or 20 weeks thereafter, the subject has received one or more 8 mg maintenance doses of VEGF receptor fusion protein; and then the method comprises administering to the subject one or more 8 mg maintenance doses of VEGF receptor fusion protein 12 weeks, 16 weeks, or 20 weeks from the last maintenance dose of VEGF receptor fusion protein and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; in, (i) The HDq12 dosing regimen includes: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 weeks after the immediately preceding dose; (ii) The HDq16 dosing regimen comprises: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered approximately 16 weeks after the immediately preceding dose; and (iii) The HDq20 dosing regimen comprises: A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 20 weeks after the immediately preceding dose.

36. A method for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, wherein the subject has received a dosing regimen for treating or preventing the nAMD, wherein the dosing regimen requires A single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by one or more second doses of about 2 mg of the VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of the VEGF receptor fusion protein ； wherein each second dose is administered about 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 8 weeks after the immediately preceding dose; and wherein the subject is at any stage of the 2 mg VEGF receptor fusion protein dosing regimen, The method comprises administering to the eye of the subject 8mg dose of VEGF receptor fusion protein, The subject is evaluated at about 8 weeks, 10 weeks, or 12 weeks after the administration, and if the treating physician determines that dosing every 12 weeks or every 16 weeks is appropriate, the subject continues to be administered 8 mg of the VEGF receptor fusion protein every 12 weeks or every 16 weeks; or The subject is evaluated within about 8 weeks, 10 weeks, or 12 weeks after the administration, and if the treating physician determines that dosing every 12 weeks is appropriate, another 8 mg dose of VEGF receptor fusion protein is administered, the subject is re-evaluated within about 12 weeks, and if the treating physician determines that dosing every 16 weeks is appropriate, the subject continues to be administered 8 mg of VEGF receptor fusion protein every 16 weeks.

37. The method of any one of claims 1 to 36, wherein the subject has received a dosing regimen for treating or preventing neovascular age-related macular degeneration: A single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by two second doses of about 2 mg of the VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of the VEGF receptor fusion protein; wherein each second dose is administered about 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 8 weeks after the immediately preceding dose.

38. The method of any one of claims 1 to 37 for treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 weeks to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 weeks to 16 weeks after the immediately preceding dose; The method further comprises extending the interval between the third doses after receiving one or more of the third doses about 12 weeks or 16 weeks after the immediately preceding dose. ●Extended from 12 weeks to 16 weeks; ● From 12 weeks to 20 weeks; or ● Extended from 16 weeks to 20 weeks, This is after the immediately preceding dose.

39. The method of claim 38, wherein the third dosage interval is adjusted at about 48 weeks or 60 weeks after initiation of treatment.

40. The method of any one of claims 38 to 39, wherein prior to said extending, said subject exhibits (a) BCVA loss < 5 letters; and / or (b) CRT < 300 or 320 μm.

41. The method of any one of claims 38 to 40, further comprising evaluating the subject's BVCA and / or CRT, and if the subject exhibits (a) BCVA loss < 5 letters; and / or (b) CRT < 300 or 320 μm, The third dosage interval is then extended.

42. The method of any one of claims 1 to 41 for treating or preventing nAMD in a subject in need thereof, the method comprising administering to the subject an initial single dose of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12 weeks to 16 weeks or 20 weeks after the immediately preceding dose, The method further comprises shortening the interval between the third doses after receiving one or more of the third doses about 12 weeks, 16 weeks, or 20 weeks after the immediately preceding dose. ●From 12 weeks to 8 weeks; ●From 16 weeks to 12 weeks; ●Shortened from 16 weeks to 8 weeks, ●Shortened from 20 weeks to 8 weeks, ●From 20 weeks to 12 weeks, or ●Shortened from 20 weeks to 16 weeks.

43. The method of claim 42, wherein prior to said shortening, said subject exhibits (a) BCVA loss >10 letters from baseline; and / or (b) CRT increase relative to baseline >50 μm.

44. The method of any one of claims 42 to 43, further comprising evaluating the subject's BVCA and / or CRT, and if the subject exhibits (a) BCVA loss >10 letters from baseline; and / or (b) CRT increase >50 μm relative to baseline, Then the third dosage interval is shortened.

45. The method of any one of claims 1 to 44, wherein if the patient is treated at week 16 or week 20 after initiation of treatment (a) a BCVA (ETDRS) loss of greater than 5 letters relative to the BCVA observed approximately 12 weeks after the initiation of treatment; (b) an increase in CRT of greater than 25 microns relative to the CRT observed about 12 weeks after initiation of treatment; and / or (c) new foveal neovascularization or foveal hemorrhage; The interval between the third dose is shortened from 12 weeks or 16 weeks to 8 weeks; or if the interval between the third dose is shortened from 12 weeks or 16 weeks to 8 weeks after the start of treatment (a) a BCVA (ETDRS) loss of greater than 5 letters relative to the BCVA observed approximately 12 weeks after the initiation of treatment; (b) an increase in CRT of greater than 25 microns relative to the CRT observed about 12 weeks after initiation of treatment; and / or (c) new foveal neovascularization or foveal hemorrhage; Then, the interval between the third dose is shortened from 16 weeks to 12 weeks.

46. ​​A method as described in any one of claims 1 to 45, for treating or preventing neovascular age-related macular degeneration in a subject in need thereof, the method comprising administering to the subject's eye three doses of about 8 mg of a VEGF receptor fusion protein in the form of a formulation comprising about 114.3 mg / ml VEGF receptor fusion protein at intervals of once every 4 weeks; and after the three doses, administering one or more doses of the VEGF receptor fusion protein at intervals extending to 12 weeks, 16 weeks or 20 weeks.

47. A method for treating or preventing nAMD in a subject in need thereof, the method comprising administering to an eye of the subject A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by 2 second doses of about 8 mg or more of the VEGF receptor fusion protein, wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and, after said dose, a) determining whether the subject meets at least one criterion for reducing or extending one or more intervals between doses of VEGF receptor fusion protein by 2 weeks, 3 weeks, 4 weeks, or 2-4 weeks; and b) if said determination is made, administering further doses of the VEGF receptor fusion protein at said reduced or extended dose intervals, The criteria for reducing the interval include:

1. BCVA loss > 5 letters; 2. Increase in central retinal thickness (CRT) > 25 μm; 3. New foveal hemorrhage; and / or 4. New foveal neovascularization, The criteria for extending the interval include:

1. BCVA loss < 5 letters, 2. There is no liquid in the central partition; 3. No new foveal hemorrhage; and / or 4. No foveal neovascularization.

48. The method of claim 47, wherein the criteria for extending the interval are include:

1. BCVA loss < 5 letters from week 12; 2. No fluid in the central partition on OCT, and 3. No new foveal hemorrhage or foveal neovascularization and / or wherein the criteria for reducing said interval include both of the following:

1. BCVA loss > 5 letters since week 12, and 2. Increase in central retinal thickness (CRT) >25 μm or new foveal hemorrhage or new foveal neovascularization from week 12 onwards.

49. The method of claim 48, wherein if the criteria are met, the interval is extended to 12 weeks, 16 weeks or 20 weeks.

50. The method of any one of claims 1 to 49 for treating or preventing neovascular age-related macular degeneration (nAMD), wherein the subject has been pre-treated with one or more 2 mg doses of a VEGF receptor fusion protein, the method comprising administering to the subject's eye A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

51. A method for treating or preventing an angiogenic eye disease in a subject in need thereof, the method comprising administering to the eye of the subject (1) a single initial dose of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 8 weeks after the immediately preceding dose; or (2) Administering a dose of 8 mg or more of VEGF receptor fusion protein once or more approximately every 4 weeks.

52. The method of claim 51, wherein the angiogenic eye disease is nAMD.

53. The method of any one of claims 51 to 52, wherein one or more second doses is 2, 3 or 4 second doses and / or about 2-4 weeks is about 4 weeks.

54. The method of any one of claims 1 to 53, wherein a subject has any one or more of the following Infection in or around the eye; Active intraocular inflammation; and / or Hypersensitivity reactions; Excluded from treatment or prevention.

55. The method of any one of claims 1 to 54, further comprising: The following steps are involved: The subjects were evaluated for the following items: Infection in or around the eye; Active intraocular inflammation; and / or Hypersensitivity reactions; And if any one or more of the above are found in the subject, the subject is excluded from treatment or prevention.

56. The method of any one of claims 1 to 55, further comprising monitoring the subject for conjunctival hemorrhage, cataract, vitreous detachment, vitreous floaters, corneal epithelial defects, and / or increased intraocular pressure during the treatment or prevention period.

57. The method of any one of claims 1 to 56, comprising, prior to any administration, providing A single-dose glass vial with a protective plastic cap and stopper containing approximately 70 microliters of an aqueous formulation containing 8 mg of VEGF receptor fusion protein; · One No. 18 x 1 1 / 2 Inch, 5 micron filter needle, which includes a bevel; ·One No. 30 x 1 / 2 Inch injection needles; and A 1-mL Luer-lock syringe with a 70-μL capacity graduation; packaged together; then (1) visually inspecting the aqueous formulation in which the VEGF receptor fusion protein is provided and using another vial of the aqueous formulation if particulates, turbidity, or discoloration is visible; (2) Remove the protective plastic cap from the vial ； and (3) clean the top of the vial with an alcohol wipe; then Use aseptic technique: (4) Take out the No. 18 x 1 from their packaging 1 / 2 Inch, 5 micron filter needle and said 1 mL syringe; (5) attaching the filter needle to the syringe by screwing the filter needle onto the Luer lock syringe tip; (6) Push the filter needle into the center of the vial stopper until the needle is fully inserted into the vial and the tip touches the bottom or bottom edge of the vial; (7) Draw all the contents of the VEGF receptor fusion protein vial into the syringe, keeping the vial in an upright position and slightly tilted, while ensuring that the inclined surface of the filter needle is immersed in the liquid; (8) continuing to tilt the vial during extraction to keep the bevel of the filter needle immersed in the formulation; (9) pulling the plunger rod back sufficiently when emptying the vial to completely empty the filter needle; (10) removing the filter needle from the syringe and disposing of the filter needle; (11) Take out the No. 30 x from its packaging 1 / 2 inch injection needle and attaching the injection needle to the syringe by firmly screwing the injection needle onto the Luer lock syringe tip; (12) holding the syringe with the needle facing upward, and checking the syringe for bubbles, wherein if there are bubbles, tapping the syringe gently with a finger until the bubbles rise to the top; and (13) Slowly depress the plunger so that the plunger tip is aligned with the line marked 70 μl on the syringe.

58. The method of any one of claims 1 to 57, wherein the VEGF inhibitor is a VEGF receptor fusion protein and the injection of the VEGF receptor fusion protein is performed under controlled aseptic conditions, including surgical hand disinfection and the use of sterile gloves, sterile drapes and sterile eyelid speculum (or equivalent), and an anesthetic and a topical broad-spectrum microbicide are administered prior to the injection.

59. The method of any one of claims 1 to 58, wherein the subject has received a dosing regimen for the treatment or prevention of nAMD requiring A single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by two second doses of about 2 mg of the VEGF receptor fusion protein, followed by one or more third doses of about 2 mg of the VEGF receptor fusion protein; wherein each second dose is administered about 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 8 weeks after the immediately preceding dose; The subject is in any stage (initial dose, second dose or third dose) of the 2 mg VEGF receptor fusion protein dosing regimen.

60. The method of any one of claims 1 to 59, wherein - one or more second doses as 2 second doses; -2 to 4 weeks is about 4 weeks; -12-20 weeks is about 12 weeks; -12-20 weeks is about 16 weeks; -12-20 weeks is about 20 weeks; -12-20 weeks is about 12-16 weeks; - 8-16 weeks is about 12 weeks; -8-16 weeks is about 16 weeks; -8-16 weeks is about 12-16 weeks; - 2 to 4 weeks is about 4 weeks and one or more second doses is 2 second doses; - 12-20 weeks is about 12 weeks and one or more second doses is 2 second doses; - 12-20 weeks is about 16 weeks and one or more second doses is 2 second doses; - 12-20 weeks is about 20 weeks and one or more second doses is 2 second doses; - 12-20 weeks is about 12-16 weeks and one or more second doses is 2 second doses; - 2 to 4 weeks is about 4 weeks and the one or more second doses is 2 second doses, and the VEGF receptor fusion protein is aflibercept; - 12-20 weeks is about 12 weeks and the one or more second doses are 2 second doses, and the VEGF receptor fusion protein is aflibercept; - 12-20 weeks is about 16 weeks and the one or more second doses is 2 second doses, and the VEGF receptor fusion protein is aflibercept; - 12-20 weeks is about 20 weeks and the one or more second doses is 2 second doses, and the VEGF receptor fusion protein is aflibercept; and / or -12-20 weeks is about 12-16 weeks and one or more second doses is 2 second doses, and the VEGF receptor fusion protein is aflibercept.

61. The method of any one of claims 1 to 60, wherein the VEGF receptor fusion protein: (i) comprising two polypeptides, the 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 130-231 of SEQ ID NO:2; and (3) a multimeric component comprising amino acids 232-457 of SEQ ID NO:2; (ii) comprising two polypeptides comprising the immunoglobulin-like (Ig) domain 2 of VEGFR1, the Ig domain 3 of VEGFR2, and a multimerization component; (iii) comprising two polypeptides comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, Ig domain 3 of VEGFR2, Ig ​​domain 4 of VEGFR2, and a multimerization component; (iv) comprising two VEGFR1R2-FcΔC1(a) polypeptides encoded by the nucleic acid sequence of SEQ ID NO: 1; or (v) selected from the group consisting of: aflibercept and conbercept.

62. The method of claim 61, wherein the VEGF receptor fusion protein comprises amino acids 27-457 of the amino acid sequence shown in SEQ ID NO:

2.

63. The method of any one of claims 1 to 62, wherein the 8 mg VEGF receptor fusion protein is in an aqueous pharmaceutical formulation selected from the group consisting of A-KKKK.

64. The method of any one of claims 1 to 63, wherein 8 mg of VEGF receptor fusion protein is administered in the form of an aqueous pharmaceutical formulation comprising 114.3 mg / ml of VEGF receptor fusion protein.

65. The method of any one of claims 1 to 64, wherein the VEGF receptor fusion protein is administered intravitreally from a syringe or a prefilled syringe.

66. The method of claim 65, wherein the syringe or pre-filled syringe is glass or plastic, and / or is sterile.

67. The method of any one of claims 1 to 66, wherein the VEGF receptor fusion protein is prepared using 30× 1 / 2 Intravitreal injection using a 1.5-inch sterile needle.

68. The method of any one of claims 1 to 67, wherein the subject has previously received one or more doses of 2 mg VEGF receptor fusion protein.

69. The method of any one of claims 1 to 68, wherein one or more additional doses are administered.

70. The method of any one of claims 1 to 69, wherein 2 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising 40 mg / ml of VEGF receptor fusion protein.

71. The method of claim 70, wherein 2 mg of VEGF receptor fusion protein is in a pharmaceutical formulation comprising: 40 mg / ml VEGF receptor fusion protein, 10 mM sodium phosphate, 40 mM NaCl, 0.03% polysorbate 20 and 5% sucrose, pH 6.

2.

72. The method of any one of claims 1 to 71, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising a sugar or a polyol.

73. The method of any one of claims 1 to 72, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising sucrose.

74. The method of any one of claims 1 to 73, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation at a pH of about 5.

8.

75. The method of any one of claims 1 to 74, wherein 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 103-126 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine.

76. The method of any one of claims 1 to 75, wherein 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 114.3 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine.

77. The method of any one of claims 1 to 76, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation, wherein the aflibercept has less than about 3.5% high molecular weight species immediately after production and purification and / or has less than or equal to about 6% high molecular weight species after storage for about 24 months at about 2-8°C.

78. The method of any one of claims 1 to 77, wherein the >8 mg VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein; About 10mM-100mM L-arginine; sucrose; Histidine-based buffers; and Surfactants; wherein the pH of the formulation is about 5.0 to about 6.8; wherein the VEGF receptor fusion protein has less than about 3.5% high molecular weight species immediately after production and purification and / or has less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months.

79. The method of claims 1 to 78, wherein 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising ●>100mg / 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; 10mM L-arginine; pH 5.8; 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; 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; 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; 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; ●>100 mg / ml aflibercept, histidine-based buffer, and L-arginine; >100 mg / ml aflibercept, about pH 5.8, wherein the formulation forms <3% HMW aggregates after incubation at 5°C for 2 months; About 114.3 mg / mL aflibercept; 10 mM-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 based buffer, 5% sucrose, 0.03% polysorbate-20, 50 mM L-arginine, pH 5.

8.

80. The method of any one of claims 1 to 79, wherein the 8 mg VEGF receptor fusion protein is in 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, l, 72μl, 73μl, 74μl, 75μl, 76μl, 77μl, 78μl, 79μl, 80μl, 81μl, 82μl, 83μl, 84μl, 85μl, 86μl l, 87μl, 88μl, 89μl, 90μl, 91μl, 92μl, 93μl, 94μl, 95μl, 96μl, 97μl, 98μl, 99μl or 100μl.

81. The method of claim 80, wherein the VEGF receptor fusion protein is administered in about 70+4 or 5 microliters.

82. The method of any one of claims 1 to 81, comprising administering the VEGF receptor fusion protein to both eyes of the subject.

83. The method of any one of claims 1 to 82, wherein the subject achieves and / or maintains one or more of: ●Best corrected visual acuity (BCVA) improved by >5, >10, >15, or >20 letters; No decrease in best corrected visual acuity (BCVA); ● Eliminate retinal fluid; ● Elimination of intraretinal fluid (IRF) and / or subretinal fluid; ●The total lesion choroidal neovascularization (CNV) area is reduced; ●Loss or reduction of intraretinal fluid; ●Loss or decrease of subretinal fluid; ●Reduction in central subretinal thickness (CST); ● Improvement of vision-related quality of life; ● Lack of adverse events (AEs) and / or serious AEs (SAEs) that emerged during the treatment period; An ETDRS letter score of at least 69 (approximately 20 / 40 Snellen equivalent); ● An increase in BCVA of ≥5, ≥10, ≥15, or ≥20 letters as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart, or no loss during the course of treatment; The increase in mean BCVA over a 12-week period; ●No intraretinal fluid (IRF) and subretinal fluid; ●Reduction in size of choroidal neovascularization (CNV); ●Reduction in total lesion CNV area compared to baseline; Loss of IRF and / or SRF; ●Reduction in central subretinal thickness (CST); Improvements in vision-related quality of life as measured by the National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25); ● Lack of adverse events (AEs) and / or serious AEs (SAEs) that emerged during the treatment period; Efficacy and / or safety in subjects with nAMD that is similar to that of aflibercept administered intravitreally at 2 mg approximately every 4 weeks for the first 3 months, followed by approximately once every 8 weeks or once every 2 months, wherein efficacy is measured as an increase in BCVA and / or a decrease in central retinal thickness, and wherein safety is measured as the incidence of adverse events such as increased blood pressure, increased intraocular pressure, visual disturbances, vitreous floaters, vitreous detachment, iris neovascularization, and / or vitreous hemorrhage; ●No anti-drug antibodies were detected during treatment; Improvement in best corrected visual acuity (BVCA) from the start of treatment (baseline) to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, Week 48, Week 52, Week 56, or Week 60; ● An increase in BCVA of ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or >7 letters as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or Snellen equivalent; ●BCVA improvement of approximately 2 or 3 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 3 letters (ETDRS or Snellen equivalent) on HDq16 at 4 weeks after the start of treatment; ●BCVA improvement eight weeks after the start of treatment of approximately 5 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 4 or 5 letters (ETDRS or Snellen equivalent) on HDq16; ●BCVA improvement of approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq16 at 12 weeks after the start of treatment; ●BCVA improvement 16 weeks after the start of treatment of approximately 6 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq16; ●BCVA improvement 20 weeks after the start of treatment of approximately 6 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16; An improvement in BCVA of approximately 5 or 6 letters (ETDRS or Snellen equivalent) at 24 weeks after the start of treatment on HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq16; ●BCVA improvement 28 weeks after the start of treatment of approximately 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16; An improvement in BCVA of approximately 6 or 7 letters (ETDRS or Snellen equivalent) at 32 weeks after the start of treatment on HDq12 or approximately 7 letters (ETDRS or Snellen equivalent) on HDq16; ●BCVA improvement of approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16 at 36 weeks after the start of treatment; An improvement in BCVA 40 weeks after the start of treatment of approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq16; Improvement in BCVA 44 weeks after initiation of treatment of approximately 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq16; An improvement in BCVA of approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16 at 48 weeks after the start of treatment; ●BCVA improvement at 52 weeks after the start of treatment of approximately 7 or 8 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq16; ●BCVA improvement of approximately 6 or 7 letters (ETDRS or Snellen equivalent) at 56 weeks after the start of treatment on HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq16; An improvement in BCVA of approximately 6 or 7 letters (ETDRS or Snellen equivalent) at 60 weeks after the start of treatment on HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq16; An improvement in BCVA of about 6 or 7 letters (ETDRS or Snellen equivalent) on HDq12 or about 6 or 7 letters (ETDRS or Snellen equivalent) on HDq16 from about 48 to about 60 weeks after the start of treatment; - An improvement in BCVA of approximately 5, 10, or 15 letters (ETDRS or Snellen equivalent) at 60 weeks after the start of treatment on the HDq12 regimen or the HDq16 regimen; An improvement in BCVA at approximately Week 8, 9, 10, 11, or 12 after the start of treatment that is maintained thereafter during the treatment regimen (within approximately +1 or +2 ETDRS letters or Snellen equivalents); BCVA of approximately 63 letters (ETDRS or Snellen equivalent) 4 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 63 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 65 letters (ETDRS or Snellen equivalent) at 8 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 65 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 letters (ETDRS or Snellen equivalent) 12 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 letters (ETDRS or Snellen equivalent) 16 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 letters (ETDRS or Snellen equivalent) at 20 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 letters (ETDRS or Snellen equivalent) at 24 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 67 letters (ETDRS or Snellen equivalent) on the HDq12 regimen or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen 28 weeks after the start of treatment; BCVA of approximately 67 letters (ETDRS or Snellen equivalent) at 32 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 67 letters (ETDRS or Snellen equivalent) at 36 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 67 letters (ETDRS or Snellen equivalent) on the HDq12 regimen or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen 40 weeks after the start of treatment; BCVA of approximately 68 letters (ETDRS or Snellen equivalent) on the HDq12 regimen or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen 44 weeks after the start of treatment; BCVA of approximately 67 letters (ETDRS or Snellen equivalent) on the HDq12 regimen or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen 48 weeks after the start of treatment; BCVA of approximately 67 or 68 letters (ETDRS or Snellen equivalent) at 52 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 or 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 or 67 letters (ETDRS or Snellen equivalent) at 56 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 or 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA of approximately 66 or 67 letters (ETDRS or Snellen equivalent) at 60 weeks after the start of treatment on the HDq12 regimen; or BCVA of approximately 66 or 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; - BCVA of about 66 to about 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen between about 48 weeks and about 60 weeks after the initiation of treatment; or BCVA of about 66 to about 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; The retina has no fluid in the central subregions (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]); ●No subretinal pigment epithelial fluid; ●No fluid leakage based on fluorescein angiography (FA); A decrease in central retinal thickness (CRT) of at least about 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 microns; The change in central retinal thickness 4 weeks after the start of treatment is about -120 or -121 or -122 or -122.4 or -120.2 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -126 or -127 or -126.6 or -126.3 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 8 weeks after the start of treatment is about -132, -133, -134, -135, or -136, or -136.2, or -132.8 microns (+ about 10, 11, or 12 microns) on the HDq12 regimen; or about -139, -140, or -139.5, or -139.6 microns (+ about 10, 11, or 12 microns) on the HDq16 regimen; The change in central retinal thickness 12 weeks after the start of treatment is about -136, -137, -138, -139, -140, -141, -140.9, or -136.6 microns (+ about 10, 11, or 12 microns) on the HDq12 regimen; or about -144, -143, or -143.5 microns (+ about 10, 11, or 12 microns) on the HDq16 regimen; The change in central retinal thickness 16 weeks after the start of treatment is about -120 or -121 or -122 or -123 or -124 or -123.4 or -120.1 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -132 or -133 or -132.1 or -133.1 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 20 weeks after the start of treatment is about -110 or -111 or -112 or -113 or -114 or -113.6 or -110.9 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -115 or -116 or -117 or -118 or -115.8 or -117.7 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 24 weeks after the start of treatment is about -134, -135, -136 or -137 or -138 or -137.6 or -134.9 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -105 or -106 or -107 or -108 or -105.3 or -107.8 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 28 weeks after the start of treatment is about -130, -131 or -132 or -133 or -134 or -133.7 or -130.7 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -144 or -145 or -146 or -147 or -148 or -144.7 or -147.2 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 32 weeks after the start of treatment is about -118 or -19 or -120 or -121 or -120.4 or -118.1 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -141 or -142 or -143 or -144 or -141.5 or -144 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 36 weeks after the start of treatment is about -142 or -143 or -144 or -144.2 or -142.2 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -126 or -127, -128 or -129 or -130 or -131 or -126.4 or -130.5 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; Change in central retinal thickness 40 weeks after initiation of treatment of approximately -131, -132, -133, -134, -133.8, or -131.2 microns (+ approximately 10, 11, or 12 microns) on the HDq12 regimen; or approximately -127, -128, or -127.5 microns (+ approximately 10, 11, or 12 microns) on the HDq16 regimen; The change in central retinal thickness 44 weeks after the start of treatment is about -120 or -121 or -122 or -123 or -124 or -125 or -124.7 or -120.3 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -143, -144 or -145 or -144.8 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; The change in central retinal thickness 48 weeks after the start of treatment is about -142 or -143 or -144 or -144.4 or -142.3 microns (+ about 10, 11 or 12 microns) on the HDq12 regimen; or about -143 or -144 or -145 or -146 or -147 or -148 or -143.8 or -147.1 microns (+ about 10, 11 or 12 microns) on the HDq16 regimen; ● Change in central retinal thickness 52 weeks after initiation of treatment of approximately -143.2 microns (+ approximately 10, 11, or 12 microns) on the HDq12 regimen; or approximately -139.6 microns (+ approximately 10, 11, or 12 microns) on the HDq16 regimen; ● Change in central retinal thickness 56 weeks after initiation of treatment of approximately -136.3 microns (+ approximately 10, 11, or 12 microns) on the HDq12 regimen; or approximately -137.5 microns (+ approximately 10, 11, or 12 microns) on the HDq16 regimen; ● Change in central retinal thickness 60 weeks after initiation of treatment of approximately -151.8 microns (+ approximately 10, 11, or 12 microns) on the HDq12 regimen; or approximately -148.8 microns (+ approximately 10, 11, or 12 microns) on the HDq16 regimen; Central retinal thickness at week 4 after initiation of treatment was approximately 248.2 microns on the HDq12 regimen; or approximately 244.1 microns on the HDq16 regimen; Central retinal thickness at week 8 after initiation of treatment was approximately 234.4 microns on the HDq12 regimen; or approximately 231.2 microns on the HDq16 regimen; Central retinal thickness at week 12 after initiation of treatment was approximately 229.7 microns on the HDq12 regimen; or approximately 226.7 microns on the HDq16 regimen; Central retinal thickness at week 16 after initiation of treatment was approximately 247.2 microns on the HDq12 regimen; or approximately 238.6 microns on the HDq16 regimen; Central retinal thickness at week 20 after initiation of treatment: approximately 257 microns for the HDq12 regimen; or approximately 254.9 microns for the HDq16 regimen; Central retinal thickness at week 24 after initiation of treatment was approximately 233 microns on the HDq12 regimen; or approximately 265.4 microns on the HDq16 regimen; Central retinal thickness at week 28 after initiation of treatment was approximately 236.9 microns on the HDq12 regimen; or approximately 226 microns on the HDq16 regimen; Central retinal thickness at week 32 after initiation of treatment was approximately 250.2 microns on the HDq12 regimen; or approximately 229.2 microns on the HDq16 regimen; Central retinal thickness at week 36 after initiation of treatment was approximately 226.4 microns on the HDq12 regimen; or approximately 244.3 microns on the HDq16 regimen; Central retinal thickness at week 40 after initiation of treatment was approximately 236.8 microns on the HDq12 regimen; or approximately 243.7 microns on the HDq16 regimen; Central retinal thickness at week 44 after initiation of treatment was approximately 245.9 microns on the HDq12 regimen; or approximately 227.7 microns on the HDq16 regimen; Central retinal thickness at week 48 after initiation of treatment: approximately 226.2 microns on the HDq12 regimen; or approximately 226.9 microns on the HDq16 regimen; Central retinal thickness at week 52 after initiation of treatment was approximately 227.4 microns on the HDq12 regimen; or approximately 231.1 microns on the HDq16 regimen; Central retinal thickness at week 56 after initiation of treatment was approximately 234.3 microns on the HDq12 regimen; or approximately 233.2 microns on the HDq16 regimen; Central retinal thickness at week 60 after initiation of treatment was approximately 218.8 microns on the HDq12 regimen; or approximately 221.9 microns on the HDq16 regimen; ● CRT at about 4, 5, 6, 7, or 8 weeks after initiation of treatment or a decrease in CRT at 4, 5, 6, 7, or 8 weeks after initiation of treatment that is thereafter maintained (within about +10, +11, or +12 microns) during the treatment regimen; ● Free aflibercept concentration in plasma was approximately 0.0409 (+0.0605) or 0 mg / L (or <0.0156 mg / L) approximately 4 hours after initiation of HDq12 or HDq16 treatment; ● Free aflibercept concentrations in plasma were approximately 0.05 (+3.78), 0.0973 (+0.102), or 0.0672 mg / L approximately 8 hours after initiation of HDq12 or HDq16 treatment; On approximately day 2 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.11 (+2.21), 0.146 (+0.110), or 0.0903 mg / L; ● On approximately day 3 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.11 (+2.06), 0.137 (+0.0947), or 0.112 mg / L; ● On approximately day 5 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.08 (+1.86), 0.0933 (+0.0481), or 0.0854 mg / L; ● On approximately day 8 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.07 (+1.75), 0.0794 (+0.0413), or 0.0682 mg / L; ● On approximately day 15 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.04 (+1.76), 0.0435 (+0.0199), or 0.0385 mg / L; On approximately day 22 after initiation of HDq12 or HDq16 treatment, the free aflibercept concentration in plasma was approximately 0.02 (+1.76), 0.0213 (+0.0148), or 0.0232 mg / L; ● Free aflibercept concentration in plasma was approximately 0.00766 (+0.00958) or 0 mg / L (or <0.0156 mg / L) on approximately day 29 after initiation of HDq12 or HDq16 treatment; ● The adjusted conjugated aflibercept concentration in plasma was approximately 0.00 mg / L approximately 4 hours after dosing following initiation of HDq12 or HDq16 treatment; ● The adjusted conjugated aflibercept concentration in plasma was approximately 0.00 mg / L approximately 8 hours after dosing following initiation of HDq12 or HDq16 treatment; The adjusted conjugated aflibercept concentration in plasma was approximately 0.06 (+3.50) or 0.124 (+0.186) mg / L on approximately day 2 after initiation of HDq12 or HDq16 treatment; The adjusted conjugated aflibercept concentration in plasma was approximately 0.13 (+2.07) or 0.173 (+0.155) mg / L on approximately day 3 after initiation of HDq12 or HDq16 treatment; The adjusted conjugated aflibercept concentration in plasma was approximately 0.18 (+1.88) or 0.223 (+0.157) mg / L on approximately day 5 after initiation of HDq12 or HDq16 treatment; ● On approximately day 8 after initiation of HDq12 or HDq16 treatment, the adjusted conjugated aflibercept concentration in plasma was approximately 0.31 (+1.56) or 0.334 (+0.135) mg / L; The adjusted conjugated aflibercept concentration in plasma was approximately 0.37 (+1.50) or 0.393 (+0.130) mg / L on approximately day 15 after initiation of HDq12 or HDq16 treatment; The adjusted conjugated aflibercept concentration in plasma was approximately 0.25 (+3.00) or 0.335 (+0.155) mg / L on approximately day 22 after initiation of HDq12 or HDq16 treatment; ● On approximately day 29 after initiation of HDq12 or HDq16 treatment, the adjusted conjugated aflibercept concentration in plasma was approximately 0.32 (+1.39) or 0.331 (+0.0953) mg / L; ● Non-inferior BVCA compared with intravitreal aflibercept at 2 mg approximately every 4 weeks for the first 5 injections followed by 2 mg approximately every 8 weeks or every 2 months; ocular and non-ocular safety or mortality in subjects with DME was similar to that of aflibercept given intravitreally at 2 mg approximately every 4 weeks for the first 3 or 4 or 5 injections, followed by 2 mg intravitreally approximately every 8 weeks or every 2 months; Improvement in best-corrected visual acuity from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; ● An increase in best-corrected visual acuity as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or Snellen equivalent of ≥2 letters, ≥3 letters, ≥4 letters, ≥5 letters, ≥6 letters, or >7 letters from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60; The retina has no fluid in the central subregion (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) from the start of treatment to Weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60; and / or A decrease in central retinal thickness (CRT) of at least 100, 125, 130, 135, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 microns from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60.

84. The method of claim 83, wherein the dry retina lacks intraretinal fluid and / or subretinal fluid.

85. The method of claim 83, wherein dry retina is characterized by the absence of intraretinal fluid (IRF) and subretinal fluid (SRF) in the subject's eye.

86. The method of claim 83, wherein the dry retina is characterized by an absence of intraretinal fluid (IRF) and an absence of subretinal fluid (SRF) in the subject's eye after the subject has received three monthly doses of the VEGF receptor fusion protein.

87. The method of any one of claims 1 to 86, wherein ● administering to the subject 1 initial dose, 2 secondary doses, and 3 tertiary doses of >8 mg of the VEGF receptor fusion protein during the first year; ● administering to the subject 1 initial dose, 2 secondary doses, and 2 tertiary doses of >8 mg of the VEGF receptor fusion protein during the first year; or - The subject is administered 1 initial dose, 2 secondary doses, and 3 tertiary doses of the >8 mg VEGF receptor fusion protein in the first year, followed by 2-4 tertiary doses in the second year.

88. The method of any one of claims 1 to 87, wherein the dosage interval is adjusted (increased / maintained / decreased) based on visual and / or anatomical results.

89. The method of any one of claims 1 to 88, wherein 12-20 weeks is 12, 13, 14, 15, 16, 17, 18, 19 or 20 weeks; and 2-4 weeks is 2, 3, 4, 5 weeks.

90. A method of treating or preventing neovascular age-related macular degeneration (nAMD) in a subject in need thereof, the method comprising administering to the subject's eye a single initial dose of about >8 mg or more of a VEGF receptor fusion protein, Subsequently, one or more second doses of about 8 mg or more of the VEGF receptor fusion protein are administered, subsequently administering one or more third doses of about 8 mg or more of the VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and wherein each third dose is administered about 12-16 weeks after the immediately preceding dose, and wherein the VEGF receptor fusion protein is aflibercept, and The vitreous half-life of aflibercept was increased compared to the intravitreal half-life of aflibercept after intravitreal administration of 40 mg / mL aflibercept, 10 mM phosphate buffer, 5% sucrose, 0.03% polysorbate 20, 40 mM NaCl, pH 6.2-6.

3.

91. The method of any one of claims 1 to 90, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation and forms 3-20% high molecular weight (HMW) aggregates after incubation for 28 days at 37°C and <3% HMW aggregates after incubation for 2 months at 5°C.

92. The method of any one of claims 1 to 91, further comprising one or more cycles of as needed (PRN), capped PRN, or treat and extend (T&E) dosing.

93. The method of any one of claims 1 to 92, wherein the VEGF receptor fusion protein is aflibercept.

94. A kit comprising: a container comprising a VEGF receptor fusion protein; and Instructions for use of VEGF receptor fusion protein, wherein the container is a vial or a prefilled syringe, wherein the container comprises >100 mg / mL VEGF receptor fusion protein, or wherein the container comprises >114.3 mg / mL VEGF receptor fusion protein, wherein the instructions comprise instructions for administering the VEGF receptor fusion protein to a patient with nAMD, The instructions include instructions for starting treatment with 8 mg of VEGF receptor fusion protein by injection once a month (once every 4 weeks) for 3 consecutive doses, wherein the instructions include instructions that the injection interval may be extended to every 16 weeks or every 20 weeks after the initial 3 consecutive doses, and The instructions include instructions for adjusting the treatment interval based on a physician's judgment of visual and / or anatomical findings.

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