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

Abstract

The present invention relates to regimens for the treatment of angiogenic eye diseases such as DR and DME characterized by high doses of aflibercept and extended dose intervals.

Description

[0001] This application is a subsidiary of U.S. Provisional Patent Application No. 63 / 319,865, filed on March 15, 2022; U.S. Provisional Patent Application No. 63 / 404,511, filed on September 7, 2022; U.S. Provisional Patent Application No. 63 / 404,889, filed on September 8, 2022; U.S. Provisional Patent Application No. 63 / 411,589, filed on September 29, 2022; U.S. Provisional Patent Application No. 63 / 412,158, filed on September 30, 2022; and U.S. Provisional Patent Application No. 63 / 412,158, filed on November 1, 2022. No. 63 / 421,296 filed on December 22, 2022; No. 63 / 434,918 filed on December 22, 2022; No. 63 / 444,470 filed on February 9, 2023; No. 63 / 447,577 filed on February 22, 2023; and Canadian Patent Application No. 3,190,726 filed on February 22, 2023; each of which is incorporated herein by reference in its entirety.

[0002] Reference to sequence listing

[0003] This application incorporates by reference a computer-readable sequence listing in ST.26 XML format, entitled 11192WO01_Sequence, created on March 13, 2023, and containing 5,464 bytes. Technical Field

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

[0005] Diabetic retinopathy (DR) is the most common microvascular complication of diabetes. Diabetic macular edema (DME), a manifestation of diabetic retinopathy, is the leading cause of vision loss and blindness in subjects with diabetes and the most common cause of blindness in young and middle-aged adults. Without treatment, approximately half of subjects with DME will lose two or more lines of visual acuity (VA) within two years. The prevalence of clinically significant macular edema (CSME) in subjects with diabetes ranges from 2.77% to 7.6%.

[0006] Intravitreal (IVT) anti-vascular endothelial growth factor (VEGF) therapy (e.g. ) inhibit the growth and leakage of new blood vessels in the retina, and they are currently the standard of care for subjects with DME. They not only maintain visual function but also provide clinically meaningful visual gains. For most subjects, treatment of DME is long-term and lifelong to inhibit retinal edema and recurrence of choroidal neovascularization (CNV). Although currently approved IVT anti-VEGF therapies are effective and well tolerated, the need for IVT injections every 4 to 8 weeks, especially during the initial phase of treatment and maintenance, is a significant burden for physicians, subjects and caregivers. While the procedure is straightforward and relatively easy to implement, capacity issues to ensure appropriate injection frequency to achieve subject outcomes similar to those seen in the pivotal studies are an increasing challenge for individual practices and the healthcare system as a whole. In addition, high-frequency dosing results in an increased burden on subjects, such as finding transportation and missing work. A secondary impact of this burden is a reduced likelihood of non-adherence to the prescribed treatment regimen.

[0007] While the efficacy and safety of currently approved VEGF antagonist therapies in treating DME have been demonstrated, 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 improved upon, the currently available standard of care.

[0008] Increasing the molar concentration of VEGF antagonist therapeutic proteins in the dosing formulation is a potential approach to further benefit patients with chorioretinal vascular diseases, including DME. Higher doses of aflibercept administered intravenously have the potential to prolong the drug's therapeutic effect. Extending the treatment interval to every 12 or 16 weeks early after treatment initiation would reduce the number of injections during the first year of treatment. Potentially reducing the injection-related treatment burden and safety events through fewer injections could be a significant contribution to patient care and healthcare delivery.

[0009] 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 macular edema following retinal vein occlusion (RVO), diabetic macular edema (DME), and diabetic retinopathy (DR). Summary of the Invention

[0010] The present invention provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema, comprising administering one or more doses (e.g., ≥8 mg) of aflibercept such that after intravitreal injection of aflibercept, the clearance of free aflibercept from the ocular chamber is about 0.367-0.458 mL / day (e.g., 0.41 mL / day), and after the intravitreal injection of aflibercept, the amount of free aflibercept in the ocular chamber of the subject reaches the lower limit of quantification (LLOQ). The method further comprises the steps of: administering the aflibercept to the subject in a manner that is consistent with the present invention; wherein 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 is about 15 weeks after the intravitreal injection of aflibercept; and wherein 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 is about 3.5 weeks after the intravitreal injection of aflibercept; 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 species immediately after manufacture and purification and / or has less than or equal to about 6% of high molecular weight species after storage at about 2° C.-8° C. for about 24 months. In one embodiment of the invention, the aqueous pharmaceutical formulation includes an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein comprising two polypeptides, each polypeptide comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, 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. In one embodiment of the invention, the method comprises administering 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12-20 weeks (preferably 12, 16 or 20 weeks) after the immediately preceding dose.

[0011] The present invention provides a method for slowing the clearance of free aflibercept from the ocular chamber after intravitreal injection relative to the clearance of aflibercept from the ocular chamber after intravitreal injection of ≤4 mg of aflibercept, the method comprising intravitreally injecting into an eye of a subject in need thereof a single initial dose of about 8 mg or more of aflibercept, followed by one or more second doses of 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 12-20 weeks after the immediately preceding dose. In one embodiment of the invention, the clearance of free aflibercept from the ocular chamber is about 34% slower than the clearance from the ocular chamber after intravitreal injection of ≤4 mg of aflibercept, for example, wherein the clearance of free aflibercept from the ocular chamber after intravitreal injection of ≥8 mg of aflibercept is about 0.367-0.458 mL / day or 0.41 mL / day.

[0012] The present invention also provides a method for increasing the time for the amount of free aflibercept in the ocular chamber of a subject to reach the lower limit of quantification (LLOQ) after intravitreal injection of aflibercept, e.g., by more than 1.3 weeks, e.g., by more than about 6 weeks to 10 weeks, e.g., to about 15 weeks, relative to the time for the amount of free aflibercept in the ocular chamber of a subject to reach the LLOQ after intravitreal injection of about 2 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 wherein each third dose is administered about 12-20 weeks after the immediately preceding dose.

[0013] The present invention also provides a method for increasing the time it takes for free aflibercept to reach the lower limit of quantitation (LLOQ) (e.g., about 0.0156 mg / L) in the plasma of a subject following intravitreal injection of aflibercept, e.g., by more than 1.5 weeks, e.g., by about 2 weeks to about 3.5 weeks, relative to the time it takes for free aflibercept to reach the LLOQ in the plasma of a subject following intravitreal injection of about 2 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 wherein each third dose is administered about 12-20 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 aflibercept, the aqueous pharmaceutical formulation comprising one or more of the following: a histidine-based buffer, arginine (e.g., L-arginine, e.g., L-arginine HCl), a sugar or a polyol (such as sucrose), and a pH of about 5.8. In one embodiment of the invention, aflibercept has less than about 3.5% high molecular weight species immediately after production and purification and / or less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months; for example, wherein ≥ 8 mg of 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 comprising two polypeptides, each polypeptide comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, 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 less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months.

[0014] The present invention provides a method for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, improving the best corrected visual acuity (BCVA) of a subject in need thereof with DR and / or DME, or promoting retinal dryness in a subject in need thereof with DR and / or DME, the method comprising administering one or more doses of about 8 mg or more of a VEGF receptor fusion protein, preferably aflibercept, to the subject's eye once every 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, or 20 weeks, or 12-20 weeks, or 12-16 weeks, or 16-20 weeks. In an embodiment of the present invention, the method comprises

[0015] A single initial dose of about 8 mg or more of a VEGF receptor fusion protein (preferably aflibercept) is administered to the eye of the subject, 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12-20 weeks after the immediately preceding dose.

[0016] The present invention includes a method for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to an eye of the subject a single initial dose of about 8 mg or more of a VEGF receptor fusion protein (preferably aflibercept), followed by one or more 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12 weeks after the immediately preceding dose.

[0017] The present invention also includes a method for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to the subject's eye a single initial dose of about 8 mg or more of a VEGF receptor fusion protein (preferably aflibercept), followed by one or more 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 16 weeks after the immediately preceding dose.

[0018] The present invention provides a method for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) 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 (preferably aflibercept), 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 20 weeks after the immediately preceding dose.

[0019] In one embodiment of the invention, a method for treating or preventing DR and / or DME in a subject comprises administering to a subject in need thereof 8 mg of a VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 microliters), administered by intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by intravitreal injection of 8 mg of the VEGF receptor fusion protein (0.07 mL) once every 8-16 weeks (2-4 months + / - 7 days). In one embodiment of the invention, the method comprises administering 8 mg of a VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 microliters), administered by intravitreal injection every 4 weeks (approximately every 28 days + / - 7 days, once a month) for the first three doses, followed by intravitreal injection of 8 mg of the VEGF receptor fusion protein (0.07 mL) once every 12 weeks (2-4 months + / - 7 days). In one embodiment of the invention, the method comprises administering 8 mg of a VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 microliters), administered by 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 of a VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 16 weeks (2-4 months + / - 7 days). In one embodiment of the invention, the method comprises administering 8 mg of a VEGF receptor fusion protein, preferably aflibercept (0.07 mL or 70 microliters), administered by 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 of a VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 20 weeks (+ / - 7 days).

[0020] The present invention also provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof: (1) wherein the subject has received an initial 2 mg dose of a VEGF receptor fusion protein, and then the method comprises administering to the subject the initial 8 mg dose of the VEGF receptor fusion protein one month later, and administering a first 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and administering a second 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and then, thereafter, administering one or more 8 mg doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20. g maintenance dose of VEGF receptor fusion protein; (2) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein, then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein 1 month later, and administering a second 8 mg second dose of VEGF receptor fusion protein 1 month later; then, thereafter, administering one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20; (3) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein, then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein 1 month later, and administering to the subject a second 8 mg second dose of VEGF receptor fusion protein 1 month later; then, administering to the subject one or more 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20; The subject is administered a second 8 mg second dose of the VEGF receptor fusion protein, and then, every 12 weeks, 16 weeks, or 20 weeks thereafter, one or more 8 mg maintenance doses of the VEGF receptor fusion protein are administered according to the HDq12, HDq16, or HDq20 dosing schedule; (4) wherein the subject has received an initial 2 mg dose of the VEGF receptor fusion protein, and then the method comprises administering to the subject a first 8 mg maintenance dose of the VEGF receptor fusion protein one month later, and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing schedule; (5) ) wherein the subject has received an initial 2 mg dose of a VEGF receptor fusion protein and received a first 2 mg second dose of a VEGF receptor fusion protein one month later, then the method comprises administering to the subject an initial 8 mg dose of the VEGF receptor fusion protein one month later, and administering a first 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and administering a second 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and then, administering one or more 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20;(6) wherein 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 first 8 mg second dose of VEGF receptor fusion protein one month later, and administering a second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20; (7) wherein 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. wherein the subject has received a first 2 mg second dose of VEGF receptor fusion protein, 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 administering to the subject 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 schedule; (8) wherein 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 first 8 mg maintenance dose of VEGF receptor fusion protein one month later, and then administering to the subject 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 schedule; (9) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and 1 month later received a first 2 mg second dose of VEGF receptor fusion protein and 1 month later received a second 2 mg second dose of VEGF receptor fusion protein, then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein 1 month later and administering the first 8 mg second dose of VEGF receptor fusion protein 1 month thereafter; and 1 month thereafter, administering the second 8 mg second dose of VEGF receptor fusion protein receptor fusion protein dose; then, every 12 weeks, 16 weeks, or 20 weeks thereafter, administer one or more 8 mg maintenance doses of VEGF receptor fusion protein according to a dosing schedule of HDq12, HDq16, or HDq20; (10) wherein 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, then the method comprises administering to 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 weeks, 16 weeks, or 20 weeks thereafter, one or more maintenance doses of 8 mg of the VEGF receptor fusion protein are administered according to the HDq12, HDq16, or HDq20 dosing schedule; (11) wherein the subject has received an initial 2 mg dose of the VEGF receptor fusion protein and received a first 2 mg second dose of the VEGF receptor fusion protein one month later and a second 2 mg second dose of the VEGF receptor fusion protein one month later, then the method comprises administering to the subject a second 8 mg second dose of the VEGF receptor fusion protein one month later and then every 12 weeks, 16 weeks, or 20 weeks thereafter, according to the HDq12, HDq16, or HDq20 dosing schedule. (12) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and 1 month later received a first 2 mg second dose of VEGF receptor fusion protein and 1 month later received a second 2 mg second dose of VEGF receptor fusion protein, 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 a HDq12, HDq16, or HDq20 dosing schedule; (13) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and 1 month later received a first 2 mg second dose of VEGF receptor fusion protein and 1 month later received a second 2 mg second dose of VEGF receptor fusion protein, 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 a HDq12, HDq16, or HDq20 dosing schedule; The method comprises administering to the subject an initial 2 mg dose of VEGF receptor fusion protein and 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 and a third 2 mg second dose of VEGF receptor fusion protein one month later, and then administering to the subject an initial 8 mg dose of VEGF receptor fusion protein one month later and 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 then, every 12 weeks, 16 weeks, or 20 weeks thereafter, according to HDq 12 or HDq16 or HDq20 dosing regimens administering one or more 8 mg maintenance doses of VEGF receptor fusion protein; (14) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and 1 month later received a first 2 mg second dose of VEGF receptor fusion protein and 1 month later received a second 2 mg second dose of VEGF receptor fusion protein and 1 month later received a third 2 mg second dose of VEGF receptor fusion protein, then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein 1 month later and 1 month thereafter, administering a second 8 mg second dose of VEGF receptor fusion protein;Then, every 12 weeks, 16 weeks, or 20 weeks thereafter, one or more maintenance doses of 8 mg of the VEGF receptor fusion protein are administered according to a dosing schedule of HDq12, HDq16, or HDq20; (15) wherein the subject has received an initial 2 mg dose of the VEGF receptor fusion protein and received a first 2 mg second dose of the VEGF receptor fusion protein one month later and received a second 2 mg second dose of the VEGF receptor fusion protein one month later and received a third 2 mg second dose of the VEGF receptor fusion protein one month later, then the method comprises administering to the subject a second 8 mg dose one month later g second dose of VEGF receptor fusion protein; then, every 12 weeks, 16 weeks, or 20 weeks thereafter, administer one or more 8 mg maintenance doses of VEGF receptor fusion protein according to the HDq12, HDq16, or HDq20 dosing schedule; (16) wherein 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 third 2 mg second dose of VEGF receptor fusion protein 1 month later, then the method comprises The method comprises administering to the subject a first maintenance dose of 8 mg of the VEGF receptor fusion protein after 2 months and administering all other maintenance doses of 8 mg of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing schedule; (17) wherein the subject has received an initial 2 mg dose of the VEGF receptor fusion protein and received a first 2 mg second dose of the VEGF receptor fusion protein 1 month later and received a second 2 mg second dose of the VEGF receptor fusion protein 1 month later and received a third 2 mg second dose of the VEGF receptor fusion protein 1 month later. VEGF receptor fusion protein, and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method then comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months later and administering the first 8 mg second dose of VEGF receptor fusion protein one month thereafter; and administering the second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20;(18) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and 1 month later received a first 2 mg second dose of VEGF receptor fusion protein and 1 month later received a second 2 mg second dose of VEGF receptor fusion protein and 1 month later received a third 2 mg second dose of VEGF receptor fusion protein and 1 month later received a fourth 2 mg second dose of VEGF receptor fusion protein; thereafter, the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein 2 months later and 1 month thereafter, administering with a second 8 mg second dose of VEGF receptor fusion protein; then, every 12 weeks, 16 weeks, or 20 weeks thereafter, one or more 8 mg maintenance doses of VEGF receptor fusion protein are administered according to a dosing schedule of HDq12, HDq16, or HDq20; (19) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later. fusion protein, and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method then comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein two months later and thereafter administering one or more 12 or 16 weekly maintenance doses of 8 mg of VEGF receptor fusion protein according to a HDq12 or HDq16 or HDq20 dosing schedule for 12 weeks or 16 weeks thereafter; (20) wherein 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 and received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method then comprises administering to 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 weeks, 16 weeks, or 20 weeks according to a dosing schedule of HDq12, HDq16, or HDq20;(21) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter; then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months after the last maintenance dose of VEGF receptor fusion protein 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; and then administering one or more 8 mg second doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing schedule of HDq12, HDq16, or HDq20. g maintenance dose of VEGF receptor fusion protein; (22) wherein 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 third 2 mg second dose of VEGF receptor fusion protein 1 month later and received a fourth 2 mg second dose of VEGF receptor fusion protein 1 month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter; then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein 2 months after the last maintenance dose of VEGF receptor fusion protein and a 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 a dosing schedule of HDq12, HDq16, or HDq20;(23) wherein 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 third 2 mg second dose of VEGF receptor fusion protein 1 month later and received a fourth 2 mg second dose of VEGF receptor fusion protein 1 month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein 2 months after the last maintenance dose of VEGF receptor fusion protein White and thereafter for 12 weeks, 16 weeks or 20 weeks, one or more weekly maintenance doses of 8 mg of VEGF receptor fusion protein are administered according to a dosing schedule of HDq12, HDq16 or HDq20 for 12 weeks, 16 weeks or 20 weeks; or (24) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein and after 1 month received a first 2 mg second dose of VEGF receptor fusion protein and after another 1 month received a second 2 mg second dose of VEGF receptor fusion protein and after another 1 month received a third 2 mg second dose of VEGF receptor fusion protein and after another 1 month received a fourth 2 mg second dose of VEGF receptor fusion protein protein, and thereafter received one or more 2 mg maintenance doses every 8 weeks, then the method comprises administering to the subject a first 8 mg maintenance dose of the VEGF receptor fusion protein 2 months after the last VEGF receptor fusion protein maintenance dose, and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, or 16 weeks, or 20 weeks according to a HDq12, or HDq16, or HDq20 dosing regimen; wherein (i) the HDq12 dosing regimen comprises: a single initial dose of the VEGF receptor fusion protein of about 8 mg or more, followed by one or more second doses of the VEGF receptor fusion protein of about 8 mg or more, followed by one or more second doses of the VEGF receptor fusion protein of about 8 mg or more, wherein each second dose is administered about 2 to 4 weeks (preferably 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 a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of a VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 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 a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of a VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 20 weeks after the immediately preceding dose - preferably, wherein the VEGF receptor fusion protein is aflibercept.;

[0021] The present invention also provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the subject has received a dosing regimen for treating or preventing the condition, wherein: (a) the subject has received an initial 8 mg dose of a VEGF receptor fusion protein, and then the method comprises administering to the subject a first 8 mg second dose of the VEGF receptor fusion protein one month later and a second 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and then, thereafter, administering once or more 8 mg of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to a dosing regimen of HDq12, HDq16, or HDq20. or (b) 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 1 month later, and the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein 1 month later, and 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 a dosing schedule of HDq12, HDq16, or HDq20; or (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 1 month later. wherein 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, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein 12 weeks, 16 weeks, or 20 weeks later, and administering all other 8 mg maintenance doses of VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to a dosing schedule of HDq12, HDq16, or HDq20; or (d) wherein 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. The subject has received one or more 8 mg maintenance doses of the VEGF receptor fusion protein 12 weeks, 16 weeks, or 20 weeks after the last maintenance dose of the VEGF receptor fusion protein and a second 8 mg second dose of the VEGF receptor fusion protein 1 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 the VEGF receptor fusion protein; and then the method comprises administering to the subject one or more 8 mg maintenance doses of the VEGF receptor fusion protein 12 weeks, 16 weeks, or 20 weeks after the last maintenance dose of the VEGF receptor fusion protein and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to a dosing schedule of HDq12, HDq16, or HDq20;wherein (i) the HDq12 dosing regimen comprises: 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 a VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of a VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12 weeks after the immediately preceding dose; and (ii) the HDq16 dosing regimen comprises: 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 a VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of a VEGF receptor fusion protein. wherein each second dose is administered about 2 to 4 weeks (preferably 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 a VEGF receptor fusion protein, followed by one or more second doses of about 8 mg or more of a VEGF receptor fusion protein, followed by one or more third doses of about 8 mg or more of a VEGF receptor fusion protein; wherein each second dose is administered about 2 to 4 weeks (preferably 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. ;

[0022] The present invention also provides a method for treating or preventing an angiogenic eye disease (preferably DR and / or DME) in a subject in need thereof, the subject having received a dosing regimen for treating or preventing the condition, the dosing regimen requiring: a single initial dose of a VEGF receptor fusion protein (preferably aflibercept) of about 2 mg, followed by one or more second doses of the VEGF receptor fusion protein of about 2 mg, followed by one or more third doses of the VEGF receptor fusion protein of about 2 mg; 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 comprising administering to the subject's eye an 8 mg dose of the VEGF receptor fusion protein, evaluating the subject about 8 weeks, 10 weeks, or 12 weeks after said administration, and continuing to administer 8 mg to the subject every 12 weeks or 16 weeks if the treating physician determines that dosing every 12 weeks or every 16 weeks is appropriate. VEGF receptor fusion protein; or evaluating the subject within about 8 weeks, 10 weeks, or 12 weeks after said administration, and if the treating physician determines that dosing every 12 weeks is appropriate, administering another 8 mg dose of VEGF receptor fusion protein, re-evaluating the subject within about 12 weeks, and if the treating physician determines that dosing every 16 weeks is appropriate, continuing to administer 8 mg of VEGF receptor fusion protein to the subject every 16 weeks.

[0023] In one embodiment of the invention, the subject in the method of the invention has received a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema: a single initial dose of a VEGF receptor fusion protein (preferably aflibercept) of about 2 mg, followed by four second doses of the VEGF receptor fusion protein of about 2 mg, followed by one or more third doses of the VEGF receptor fusion protein of about 2 mg; 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.

[0024] The present invention also provides a method for treating or preventing diabetic retinopathy or diabetic macular edema in a subject in need thereof, the method comprising administering to the subject's eye a single initial dose of a VEGF receptor fusion protein (preferably aflibercept) of about 8 mg or more, followed by one or more second doses of the VEGF receptor fusion protein of about 8 mg or more, followed by one or more third doses of the VEGF receptor fusion protein of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12 or 16 weeks after the immediately preceding dose; the method further comprises, after receiving one or more third doses about 12 weeks or 16 weeks after the immediately 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 after the immediately preceding dose. For example, in one embodiment of the invention, during the treatment period, the subject exhibits (a) BCVA loss < 5 letters; and / or (b) central retinal thickness (CRT) < 300 or 320 μm. In one embodiment of the invention, the method further comprises 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.

[0025] The present invention also provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 (preferably aflibercept), 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12 weeks, 16 weeks, or 20 weeks after the immediately preceding dose; the method further comprises, after receiving one or more of the third doses about 12 weeks, 16 weeks, or 20 weeks after the immediately 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, during the treatment period, the subject exhibits (a) a BCVA loss of >10 letters relative to baseline; and / or (b) an increase in CRT of >50 μm relative to baseline. For example, in one embodiment of the invention, the method further comprises evaluating the subject's BVCA and / or CRT, and shortening the third dose interval if the subject exhibits (a) a BCVA loss of >10 letters relative to baseline; and / or (b) an increase in CRT of >50 μm relative to baseline.

[0026] In one embodiment of the invention, the interval between third doses is reduced from 12 weeks or 16 weeks to 8 weeks if (a) the BCVA (ETDRS) loss is greater than 5 letters relative to the BCVA observed approximately 12 weeks after the initiation of treatment; (b) the CRT increases by more than 25 microns relative to the CRT observed approximately 12 weeks after the initiation of treatment; and / or (c) there is new onset foveal neovascularization or foveal hemorrhage; for example, at week 16 or week 20 after the initiation of treatment; or if (a) the BCVA (ETDRS) loss is greater than 5 letters relative to the BCVA observed approximately 12 weeks after the initiation of treatment; (b) the CRT increases by more than 25 microns relative to the CRT observed approximately 12 weeks after the initiation of treatment; and / or (c) there is new onset foveal neovascularization or foveal hemorrhage; for example, at week 24 after the initiation of treatment, the interval between third doses is reduced from 16 weeks to 12 weeks.

[0027] The present invention provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 (preferably aflibercept) in the form of a formulation comprising about 114.3 mg / ml of the VEGF receptor fusion protein at an interval of once every four weeks; wherein after the three doses, one or more doses of the VEGF receptor fusion protein are administered at intervals extending to 12 weeks, 16 weeks, or 20 weeks.

[0028] The present invention includes a method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 a VEGF receptor fusion protein (preferably aflibercept), followed by two 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 (preferably 4 weeks) after the immediately preceding dose; and, after the doses, a) determining whether the subject meets at least one criterion for reducing or extending one or more intervals between doses of the VEGF receptor fusion protein by 2 weeks, 3 weeks, 4 weeks, or 2-4 weeks; and

[0029] b) if such determination is made, administering additional doses of the VEGF receptor fusion protein at such reduced or extended dose intervals, wherein criteria for extending the interval include: 1. BCVA loss <5 letters; and / or 2. CRT <300 or 320 microns; and wherein criteria for reducing the interval include: 1. BCVA loss >10 letters; 2. persistent or worsening DME; and / or 3. CRT increase >50 microns. In one embodiment of the invention, wherein criteria for extending the interval include both of the following: 1. BCVA loss <5 letters from week 12; and 2. CRT <300 or 320 microns as measured by SD-OCT; and / or wherein criteria for reducing the interval include both of the following: 1. BCVA loss >10 letters combined with persistent or worsening DME, e.g., from week 12; and 2. CRT increase >50 microns, e.g., from week 12. In one embodiment of the invention, if such criteria are met, the interval is extended to 12 weeks, 16 weeks, or 20 weeks.

[0030] The present invention provides a method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the subject having been pre-treated with one or more 2 mg doses of a VEGF receptor fusion protein (preferably aflibercept), the method comprising administering to the subject's eye a single initial 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 (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 12-20 weeks after the immediately preceding dose.

[0031] The present invention provides a method for treating or preventing angiogenic eye diseases (preferably DR and / or DME) in a subject in need thereof, the method comprising administering to the eye of the subject (1) a single initial dose of a VEGF receptor fusion protein (preferably aflibercept) of about 8 mg or more, followed by one or more second doses of the VEGF receptor fusion protein of about 8 mg or more, followed by one or more third doses of the VEGF receptor fusion protein of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks (preferably 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 one or more doses of 8 mg or more of the VEGF receptor fusion protein about every 4 weeks.

[0032] In one embodiment of the invention, subjects with certain exclusion criteria are excluded from treatment, or if the exclusion criteria are not met, are not excluded from treatment. For example, subjects with any one or more of an ocular or periocular infection; active intraocular inflammation; and / or a hypersensitivity reaction are excluded from administration of a VEGF receptor fusion protein to the eye. In one embodiment of the invention, the method of the invention further comprises the step of evaluating the subject for any one or more of the following: an ocular or periocular infection; active intraocular inflammation; and / or a hypersensitivity reaction; and if any one or more of the above is found in the subject, the subject is excluded from administration.

[0033] In one embodiment of the present invention, the subject is monitored for adverse events such as conjunctival hemorrhage, cataract, vitreous detachment, vitreous floaters, corneal epithelial defects and / or increased intraocular pressure. If such AEs are identified, the identified AEs may be treated and / or such treatment or prevention may be discontinued.

[0034] In one embodiment of the present invention, the method includes preparation prior to administration of the VEGF receptor fusion protein (preferably aflibercept). For example, the method includes providing or preparing before each administration: a single-dose glass vial with a protective plastic cap and stopper containing approximately 70 microliters of an aqueous formulation containing 8 mg of the VEGF receptor fusion protein; a filter needle, such as an 18-gauge x 1 1 1 / 2 inch, 5 micron filtered needle, including a tip and bevel; an invention needle, such as a 30 gauge x 1 / 2 inch injection needle; and a syringe, such as a 1 mL Luer lock syringe, having a graduation marking 70 microliters of capacity; packaged together; then (1) visually inspect the aqueous formulation in the vial and, if particulates, turbidity, or discoloration are visible, use another vial containing an aqueous formulation of a VEGF receptor fusion protein; (2) remove the protective plastic cap from the vial; and (3) clean the top of the vial with an alcohol wipe; then, using aseptic technique: (4) remove the 18 gauge x 1 / 2 inch syringe from their packaging; 1 1 mL syringe; (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 the needle is fully inserted into the vial and the tip touches the bottom or bottom edge of the vial; (7) withdraw the entire contents of the VEGF receptor fusion protein vial into the syringe, holding the vial in an upright position, slightly tilted, while ensuring that the bevel of the filter needle is immersed in the liquid; (8) continue to tilt the vial during withdrawal, keeping the bevel of the filter needle immersed in the formulation; (9) When emptying the vial, pull the plunger rod back sufficiently to completely empty the filter needle; (10) remove the filter needle from the syringe and dispose of it; (11) remove the 30 gauge x 1 / 2 inch injection needle from its packaging and attach it to the syringe by firmly screwing it onto the tip of the Luer lock syringe; (12) hold the syringe with the needle pointing upward and check the syringe for bubbles, wherein if bubbles are present, gently tap the syringe with your fingers until the bubbles rise to the top; and (13) slowly depress the plunger until the plunger tip is aligned with the 70 microliter mark on the syringe. In one embodiment of the present invention, injection of VEGF receptor fusion protein is performed under controlled aseptic conditions, which include surgical hand disinfection and the use of sterile gloves, sterile drapes, and sterile eye speculum (or equivalent), and an anesthetic and a topical broad-spectrum microbicide are applied prior to injection.

[0035] In one embodiment of the invention, the subject has been receiving a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema which requires: a single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by four 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; wherein the subject is at any stage (initial dose, second dose, or third dose) of the 2 mg VEGF receptor fusion protein dosing regimen.

[0036] In one embodiment of the present invention, one or more second doses are 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 are 2 second doses; 12-20 weeks is about 12 weeks and one or more second doses are 2 doses; 12-20 weeks is about 16 weeks and one or more second doses are 2 second doses; 12-20 weeks is about 20 weeks and one or more second doses are 2 doses; 12-20 weeks is about 12-16 weeks and one or more second doses are 2 second doses; 2 to 4 weeks is about 4 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 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 are 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 are 2 second doses and the VEGF receptor fusion protein is aflibercept; and / or 12-20 weeks is about 12-16 weeks and the one or more second doses are 2 second doses and the VEGF receptor fusion protein is aflibercept.

[0037] In one embodiment of the present invention, the VEGF receptor fusion protein comprises amino acids 27-457 of the amino acid sequence shown in SEQ ID NO: 2; is selected from the group consisting of aflibercept and conbercept; comprises 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 VEGFR2 component comprising amino acids 130-231 of SEQ ID NO: 2. NO:2; comprising two polypeptides comprising the immunoglobulin-like (Ig) domain 2 of VEGFR1, the Ig domain 3 of VEGFR2, and a multimerization component; comprising two polypeptides comprising the immunoglobulin-like (Ig) domain 2 of VEGFR1, the Ig domain 3 of VEGFR2, the Ig domain 4 of VEGFR2, and a multimerization component; or comprising two VEGFR1R2-FcΔC1(a) polypeptides, the two VEGFR1R2-FcΔC1(a) polypeptides being encoded by the nucleic acid sequence of SEQ ID NO: 1. In one embodiment of the present invention, the VEGF receptor fusion protein comprises or consists of amino acids 27-457 of the amino acid sequence shown in SEQ ID NO: 2.

[0038] In one embodiment of the invention, the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation selected from the group consisting of A-KKKK. In one embodiment of the invention, the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 114.3 mg / ml VEGF receptor fusion protein (preferably aflibercept).

[0039] In one embodiment of the invention, the VEGF receptor fusion protein is administered to both eyes of a subject.

[0040] In one embodiment of the invention, the VEGF receptor fusion protein (preferably aflibercept) is administered from a syringe or prefilled syringe, e.g., glass or plastic, and / or sterile; e.g., with a 30 gauge x 1 / 2 inch sterile injection needle.

[0041] In one embodiment of the invention, the subject has previously received one or more doses of 2 mg of a VEGF receptor fusion protein, such as Eylea. The subject may be administered one or more doses greater than the dose specifically mentioned.

[0042] In one embodiment of the invention, the subject who has received 2 mg of a VEGF receptor fusion protein (preferably aflibercept) has received the protein in an aqueous pharmaceutical formulation comprising 40 mg / ml of a VEGF receptor fusion protein, 10 mM sodium phosphate, 40 mM NaCl, 0.03% polysorbate 20, and 5% sucrose at a pH of 6.2.

[0043] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising >100 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine (preferably L-arginine); for example, comprising a sugar or polyol (e.g., sucrose). In one embodiment of the invention, the pH of the formulation is about 5.8. For example, the formulation may comprise about 103-126 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine; in one embodiment of the invention, about 114.3 mg / ml of VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0044] In one embodiment of the invention, 8 mg of VEGF receptor fusion protein is dissolved 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; or a volume of 100 μl; for example, administered in a volume of about 70 ± 4 or 5 microliters.

[0045] In one embodiment of the invention, the methods herein include the step of administering a VEGF receptor fusion protein (preferably aflibercept) to both eyes of the subject, such as intravitreally.

[0046] In one embodiment of the invention, the subject achieves and / or maintains one or more of the following: an improvement in the Diabetic Retinopathy Severity Scale (DRSS); an improvement in best corrected visual acuity; an improvement in best corrected visual acuity with dry retina; a BCVA of at least 69 letters; an absence of fluid in the center of the fovea; a decrease in central retinal thickness (CRT); an absence of vascular leakage as measured by fluorescein angiography (FA); an improvement in the National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score from pre-treatment baseline; an absence of fluid in the retina at the center of the fovea and in the central subregion (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [ maintenance of retinal fluid-free status (total fluid, IRF, and / or SRF in the foveal center and subregions); absence of macular edema; absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT); no weaning from HDq12, HDq16, or HDq20 treatment regimens once initiated; noninferior BVCA compared to intravitreal aflibercept at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections followed by 2 mg approximately every 8 weeks or every 2 months; an increase in BCVA (based on ETDRS letter score) of approximately 7, 8, or 9 letters from the start of treatment to week 60, where baseline BCVA was approximately 61, 62, or 63; BCVA (based on ETDRS letter score) of at least about 69 letters from the start of treatment to Week 48 or Week 60; No loss of 5, 10, 15, or 69 letters or more of BCVA after Week 12, 24, 36, 48, 60, 72, 84, 90, or 96 from the start of treatment; Improvement in BCVA (based on ETDRS letter score) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; BCVA (based on ETDRS letter score) from the start of treatment to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, or Week 48 VCA improvement; BCVA score (based on ETDRS letter score) of about 69, 70, 71, 72, or 73 between Week 48 and Week 60; a change in BCVA score (based on ETDRS letter score) of about 7, 8, or 9 from the start of treatment between Week 36 and Week 48, with the BCVA at any point between Week 36 and Week 48 being about 60 or 70; a change in BCVA score (based on ETDRS letter score) of about 7, 8, or 9 from the start of treatment between Week 48 and Week 60, with the BCVA at any point between Week 48 and Week 60 being about 69, 70, 71, 72, or 73;Gain of ≥4 letters, ≥5 letters, ≥6 letters, ≥7 letters, ≥8 letters, >9 letters, or >10 letters in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; No loss of 5, 10, or 15 letters (based on the ETDRS letter score) from the start of treatment to Week 48 or Week 60; Gain of 5, 10, or 15 letters (based on the ETDRS letter score) from the start of treatment to Week 48 or Week 60; Improvement in BCVA 4 weeks after treatment initiation of approximately 4 letters on the HDq12 regimen or 5 letters (ETDRS or Snellen equivalents) on HDq16; or about 4 or 5 letters (ETDRS or Snellen equivalents) on HDq16; BCVA improvement 8 weeks after the start of treatment is about 6 letters (ETDRS or Snellen equivalents) on HDq12; or about 5 or 6 letters (ETDRS or Snellen equivalents) on HDq16; BCVA improvement 12 weeks after the start of treatment is about 6 or 7 letters (ETDRS or Snellen equivalents) on HDq12; or about 6 letters (ETDRS or Snellen equivalents) on HDq16; The BCVA improvement at 16 weeks was approximately 6 or 7 letters (ETDRS or Snellen equivalents) with HDq12 or 7 letters (ETDRS or Snellen equivalents) with HDq16. The BCVA improvement at 20 weeks after the start of treatment was approximately 6 letters (ETDRS or Snellen equivalents) with HDq12 or 6 letters (ETDRS or Snellen equivalents) with HDq16. The BCVA improvement at 24 weeks after the start of treatment was approximately 7 letters (ETDRS or Snellen equivalents) with HDq12 or 5 or 6 letters (ETDRS or Snellen equivalents) with HDq16. ETDRS or Snellen equivalents); BCVA improvement 28 weeks after treatment initiation is approximately 7 or 8 letters (ETDRS or Snellen equivalents) when using the HDq12 regimen; or approximately 7 or 8 letters (ETDRS or Snellen equivalents) when using the HDq16 regimen; BCVA improvement 32 weeks after treatment initiation is approximately 7 letters (ETDRS or Snellen equivalents) when using the HDq12 regimen; or approximately 7 or 8 letters (ETDRS or Snellen equivalents) when using the HDq16 regimen; BCVA improvement 36 weeks after treatment initiation is 8 letters (ETDRS or Snellen equivalents) when using the HDq12 regimen;or about 6 or 7 letters (ETDRS or Snellen equivalents) on HDq16; BCVA improvement 40 weeks after treatment initiation was about 8 letters (ETDRS or Snellen equivalents) on HDq12; or about 6 or 7 letters (ETDRS or Snellen equivalents) on HDq16; BCVA improvement 44 weeks after treatment initiation was about 8 letters (ETDRS or Snellen equivalents) on HDq12; or about 7 or 8 letters (ETDRS or Snellen equivalents) on HDq16; BCVA improvement 48 weeks after treatment initiation was about 8 letters (ETDRS or Snellen equivalents) on HDq12 about 8 or 9 letters (ETDRS or Snellen equivalents) when taking the HDq12 or HDq16 regimen; or about 7 or 8 letters (ETDRS or Snellen equivalents) when taking the HDq16 regimen; an improvement in BCVA at about week 8 after the start of treatment that is maintained thereafter through at least week 48 during the treatment regimen; an improvement in BCVA score (based on ETDRS letter scores) of up to 38 letters from the start of treatment between weeks 36 and 48 when taking the HDq12 or HDq16 regimen; a BCVA of about 68 letters (ETDRS or Snellen equivalents) 4 weeks after the start of treatment when taking the HDq12 regimen; or about 66 letters (ETDRS or Snellen equivalents) when taking the HDq16 regimen BCVA at 8 weeks after the initiation of treatment was approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA of approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 12 weeks after the initiation of treatment was approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA of approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 16 weeks after the initiation of treatment was approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen. S or Snellen equivalents); or BCVA of approximately 69 letters (ETDRS or Snellen equivalents) on the HDq16 regimen; BCVA of approximately 70 letters (ETDRS or Snellen equivalents) on the HDq12 regimen 20 weeks after the initiation of treatment; or BCVA of approximately 68 letters (ETDRS or Snellen equivalents) on the HDq16 regimen; BCVA of approximately 71 letters (ETDRS or Snellen equivalents) on the HDq12 regimen 24 weeks after the initiation of treatment; or BCVA of approximately 67 letters (ETDRS or Snellen equivalents) on the HDq16 regimen;BCVA at 28 weeks after the initiation of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 32 weeks after the initiation of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 36 weeks after the initiation of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen. BCVA at 40 weeks after the initiation of treatment was approximately 72 letters (ETDRS or Snellen equivalents) on the HDq12 regimen; or approximately 69 letters (ETDRS or Snellen equivalents) on the HDq16 regimen; BCVA at 44 weeks after the initiation of treatment was approximately 72 letters (ETDRS or Snellen equivalents) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalents) on the HDq16 regimen; and BCVA at 48 weeks after the initiation of treatment was approximately 73 letters on the HDq12 regimen. letters (ETDRS or Snellen equivalents) on the HDq16 regimen; or a BCVA of approximately 70 letters (ETDRS or Snellen equivalents) on the HDq16 regimen; or a BCVA of approximately 71, 72, 73, or 74 (ETDRS or Snellen equivalents) between Weeks 36 and 48 on the HDq12 regimen; or a BCVA of approximately 69, 70, 71, 72, or 73 (ETDRS or Snellen equivalents) between Weeks 36 and 48 on the HDq16 regimen; or an improvement in BCVA of approximately 9 or 10 letters at Week 48 after the initiation of treatment on the HDq12 regimen when baseline BCVA was <73 ETDRS letters (ETDRS or Snellen equivalents); with HDq12, when baseline BCVA was approximately >73 ETDRS letters, the BCVA improvement at week 48 after the initiation of treatment was approximately 5 or 6 letters (ETDRS or Snellen equivalents); with HDq16, when baseline BCVA was approximately <73 ETDRS letters, the BCVA improvement at week 48 after the initiation of treatment was approximately 8 or 9 letters (ETDRS or Snellen equivalents); with HDq16, when baseline BCVA was approximately >73 ETDRS letters, the BCVA improvement at week 48 after the initiation of treatment was approximately 4 or 5 letters (ETDRS or Snellen equivalents);When using the HDq12 regimen, when the baseline CRT is about < about 400 microns, the BCVA improvement at week 48 after the initiation of treatment is about 7 or 8 letters (ETDRS or Snellen equivalent); when using the HDq12 regimen, when the baseline CRT is about > 400 microns, the BCVA improvement at week 48 after the initiation of treatment is about 9 or 10 letters (ETDRS or Snellen equivalent); when using the HDq16 regimen, when the baseline CRT is about < about 400 microns, the BCVA improvement at week 48 after the initiation of treatment is about 5 or 6 letters (ETDRS or Snellen equivalent). ETDRS or Snellen equivalents); an improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalents) at Week 48 after the start of treatment when the baseline CRT is approximately >400 microns on HDq16; an increase in BCVA (based on ETDRS letter score) of >5, >10, or ≥15 letters from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; severe diabetic retinopathy from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96 Improvement of ≥2 or >3 grades on the Diabetic Retinopathy Severity Scale (DRSS); improvement of ≥2 grades on the Diabetic Retinopathy Severity Scale (DRSS) from the start of treatment to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, or Week 48; absence of retinal fluid (total fluid, intraretinal fluid [IRF], and / or intraretinal fluid [IRF]) at the center of the fovea and in the central subregion as measured by optical coherence tomography (OCT) from the start of treatment to Week 12, Week 24, Week 36, Week 48, Week 60, Week 72, Week 84, Week 90, or Week 96. =Subretinal fluid [SRF]); absence of vascular leakage as measured by fluorescein angiography (FA) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; maintenance of absence of retinal fluid (total fluid, IRF, and / or SRF in the foveal center and central subregion) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; a reduction in the total area of ​​fluorescein leakage (mm2) within the ETDRS grid as measured by fluorescein angiography by approximately 12, 13, or 14 mm at Week 48 or Week 60; 2or more; no retinal fluid based on spectral domain optical coherence tomography (SD-OCT) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; no retinal fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; dry retina from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; no retinal fluid based on spectral domain optical coherence tomography (SD-OCT) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96 ) measured with no fluid in the center of the fovea; the change in central retinal thickness 4 weeks after the start of treatment was about -118 or -118.3 microns when using the HDq12 regimen; or about -124 or -125 or -124.9 or -125.5 microns when using the HDq16 regimen; the change in central retinal thickness 8 weeks after the start of treatment was about -137 or -137.4 microns when using the HDq12 regimen; or about -139 or -140 or -139.6 or -140.3 microns when using the HDq16 regimen; the change in central retinal thickness 12 weeks after the start of treatment was about -150 or -150.1 microns when using the HDq12 regimen; or about -150 or -150.1 microns when using the HDq16 regimen The change in central retinal thickness at 16 weeks after the start of treatment was about -139 or -139.4 microns when using the HDq12 regimen; or about -145 or -146 or -145.5 or -146.4 microns when using the HDq16 regimen; the change in central retinal thickness at 20 weeks after the start of treatment was about -117 or -117.1 microns when using the HDq12 regimen; or about -112 or -113 or -112.5 or -113.3 microns when using the HDq16 regimen; the change in central retinal thickness at 24 weeks after the start of treatment was about -158 or -159.4 microns when using the HDq12 regimen; or about -146 or -147.5 or -148.4 microns when using the HDq16 regimen. The change in central retinal thickness at 28 weeks after the start of treatment was about -146, -147, or -146.7 microns when using the HDq12 regimen; or about -162 or -162.3 microns when using the HDq16 regimen; the change in central retinal thickness at 32 weeks after the start of treatment was about -132 microns when using the HDq12 regimen; or about -145, -146, or -145.8 microns when using the HDq16 regimen; and the change in central retinal thickness at 36 weeks after the start of treatment was about -168 or -168.1 micron; or about -124, -125, -124.7, or -125.2 microns when using the HDq16 regimen; the change in central retinal thickness 40 weeks after the start of treatment was about -163 microns when using the HDq12 regimen; or about -122, -123, -122.5, or -123.1 microns when using the HDq16 regimen; the change in central retinal thickness 44 weeks after the start of treatment was about -147, -148, or -147.4 microns when using the HDq12 regimen; or about -164, -164.1, or -164.3 microns when using the HDq16 regimen; the change in central retinal thickness 48 weeks after the start of treatment was about -163 microns when using the HDq12 regimen; or about -122, -123, -122.5, or -123.1 microns when using the HDq16 regimen The change in central retinal thickness 60 weeks after the start of treatment was approximately -171, -172, or -171.7 microns on the HDq2 regimen; or approximately -148, -149, -148.3, or -149.4 microns on the HDq16 regimen; the change in central retinal thickness 60 weeks after the start of treatment was approximately -181.95 or -176.24 microns on the HDq12 regimen; or approximately -166.26 or -167.18 microns on the HDq16 regimen; the change in central retinal thickness between the start of treatment (week 0) and week 4 on the HDq12 regimen was approximately -118, -119, or -118.3 microns; the change in central retinal thickness between weeks 4 and 8 on the HDq12 regimen was approximately -148, -149, -148.3, or -149.4 microns. The change in central retinal thickness was approximately -19, -20, or -19.1 microns between weeks 8 and 12 on the HDq12 regimen; the change in central retinal thickness was approximately -12, -13, or -12.7 microns between weeks 20 and 24 on the HDq12 regimen; the change in central retinal thickness was approximately -40 or -41 microns between weeks 20 and 24 on the HDq12 regimen; the change in central retinal thickness was approximately -36, -37, or -36.1 microns between weeks 32 and 36 on the HDq12 regimen; the change in central retinal thickness was approximately -24, -25, or -24.3 microns between weeks 44 and 48 on the HDq12 regimen; and the change in central retinal thickness was approximately -30, -31, or -31.7 microns between weeks 48 and 49 on the HDq12 regimen. Between the start of treatment (week 0) and week 4, the change in central retinal thickness was approximately -4, -5, or -4.5 microns on the HDq16 regimen; between the start of treatment (week 0) and week 4, the change in central retinal thickness was approximately -124, -125, or -124.9 microns on the HDq16 regimen; between weeks 4 and 8, the change in central retinal thickness was approximately -14, -15, or -14.7 microns on the HDq16 regimen; between weeks 8 and 12, the change in central retinal thickness was approximately -13, -14, or -13.1 microns on the HDq16 regimen; and between weeks 24 and 28, the change in central retinal thickness was approximately -58, -59, or -58.5 microns; a change in central retinal thickness of approximately -41, -42, or -41.6 microns between Week 40 and Week 44 on the HDq16 regimen; a decrease in central retinal thickness at Week 4, 5, 6, 7, or 8 after the start of treatment that is maintained within approximately ±17, ±18, or ±19 microns during the treatment regimen and through at least Week 48 after the start of treatment; a decrease in central retinal thickness of approximately 100, 125, 150, 175, or 200 microns from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; a decrease in central retinal thickness of approximately 148-182 microns as measured by optical coherence tomography (OCT) at approximately Week 48 or 60 after the start of treatment, where the baseline a CRT of about 449, 450, 455, or 460 microns; a decrease in central retinal thickness (CRT) of at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 microns from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; and free aflibercept in plasma of about 0.149 (±0.249) mg / L at about 0.1667 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; About 0.3333 days after the first dose, the free aflibercept in the plasma was about 0.205 (± 0.250) mg / l; wherein, at baseline, free aflibercept was undetectable in the plasma, and wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 1 day after the first dose, the free aflibercept in the plasma was about 0.266 (± 0.211) mg / l; wherein, at baseline, free aflibercept was undetectable in the plasma, and wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 2 days after the first dose, the free aflibercept in the plasma was about 0.218 (± 0.145) mg / l; wherein, at baseline, free aflibercept was undetectable in the plasma. wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; at about 4 days after the first dose, the free aflibercept in the plasma is about 0.140 (± 0.0741) mg / l; wherein, at baseline, the free aflibercept in the plasma is undetectable, wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; at about 7 days after the first dose, the free aflibercept in the plasma is about 0.0767 (± 0.0436) mg / l; wherein, at baseline, the free aflibercept in the plasma is undetectable, wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; at about 14 days after the first dose, the free aflibercept in the plasma is about 0.0309 (± 0.0241) mg / l; wherein at baseline, free aflibercept was undetectable in plasma, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; at about 21 days after the first dose, free aflibercept in plasma was about 0.0171 (±0.0171) mg / l; wherein, at baseline, free aflibercept was undetectable in plasma, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; at about 28 days after the first dose, free aflibercept in plasma was about 0.00730 (±0.0113) mg / l; wherein, at baseline, free aflibercept was undetectable in plasma, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; Approximately 0.1667 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.00698 (±0.0276) mg / L; wherein, at baseline, there was approximately 0.00583 mg / L (±0.0280) of adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 0.3333 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.00731 (±0.0279) mg / L; wherein, at baseline, there was approximately 0.00583 mg / L (±0.0280) of adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 1 day after the first dose On day 1, the adjusted bound aflibercept in plasma was approximately 0.0678 (±0.0486) mg / l; wherein, at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 2 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.138 (±0.0618) mg / l; wherein, at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 4 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.25 9 (± 0.126) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 7 days after the first dose, the adjusted bound aflibercept in the plasma was approximately 0.346 (± 0.151) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 14 days after the first dose, the adjusted bound aflibercept in the plasma was approximately 0.374 (± 0.110) mg / l; wherein at baseline, there was approximately 0.0.00583 mg / l (±0.0280) adjusted bound aflibercept in the blood plasma, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; approximately 21 days after the first dose, the adjusted bound aflibercept in the plasma was approximately 0.343 (±0.128) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept in the blood plasma, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks approximately 28 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.269 (±0.149) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (±0.0280) of adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; the free aflibercept level in plasma reached a maximum level approximately 0.965 days after the first dose; the free aflibercept in plasma reached approximately 0.310 mg / l (±0.263); free aflibercept in the plasma was about 0 to about 1.08 mg / L; the maximum free aflibercept in the plasma (mg / l) per dose (mg) of aflibercept was about 0.388 (±0.0328) mg / l / mg; the adjusted bound aflibercept in the plasma reached a maximum level about 14 days after the first dose; the adjusted bound aflibercept in the plasma reached about 0.387 mg / l (±0.135) the maximum level of aflibercept in the plasma; the adjusted bound aflibercept in the plasma is about 0.137 to about 0.774 mg / L; the adjusted bound aflibercept in the plasma (mg / L) per dose (mg) of aflibercept is about 0.483 (±0.0168) mg / L / mg; there are no anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment; the National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score improves from the pre-treatment baseline; and / or there is no macular edema. For example, in one embodiment of the invention, the dry retina lacks intraretinal fluid and / or subretinal fluid; or the dry retina is characterized by the absence of intraretinal fluid (IRF) and the absence of subretinal fluid (SRF) in the subject's eye after the subject has received three monthly doses of the VEGF receptor fusion protein.

[0047] In one embodiment of the invention reference to 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56 or 60 weeks from the start of treatment is about 48 weeks or 60 weeks from the start of treatment.

[0048] In one embodiment of the present invention, the subject is administered one initial dose, two secondary doses, and three tertiary doses of a VEGF receptor fusion protein, such as aflibercept, in the first year; the subject is administered one initial dose, two secondary doses, and two tertiary doses of a VEGF receptor fusion protein, such as aflibercept, in the first year; or the subject is administered one initial dose, two secondary doses, and three tertiary doses of a VEGF receptor fusion protein, such as aflibercept, in the first year, followed by 2-4 tertiary doses in the second year.

[0049] In one embodiment of the invention, based on visual and / or anatomical findings, e.g. Figure 3 and / or Figure 4 Adjust (increase / maintain / decrease) the dosage interval based on the criteria shown in .

[0050] The present invention also provides a kit comprising: a container comprising a VEGF receptor fusion protein; and instructions for use of the VEGF receptor fusion protein, wherein the container is a vial or a prefilled syringe, wherein the container comprises ≥100 mg / mL of the VEGF receptor fusion protein, wherein the container comprises ≥114.3 mg / mL of the VEGF receptor fusion protein, wherein the instructions include instructions for administering aflibercept to a patient with DME / AMD, wherein the instructions include instructions for initiating treatment with aflibercept 8 mg with a monthly injection (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 a physician's judgment of visual and / or anatomical findings.

[0051] The present invention provides aflibercept for use in the treatment or prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the treatment or prevention comprising administering one or more doses of aflibercept at intervals and in amounts such that, following intravitreal injection of aflibercept, the clearance of free aflibercept from the ocular chamber is about 0.367-0.458 mL / day, the time to reach the lower limit of quantification (LLOQ) of the amount of free aflibercept in the ocular chamber of the subject is about 15 weeks following the intravitreal injection of aflibercept; and the time to reach the lower limit of quantification (LLOQ) of free aflibercept in the plasma of the subject is about 3.5 weeks following the intravitreal injection of aflibercept.

[0052] The present invention provides aflibercept for use in a method for slowing the clearance of free aflibercept from the ocular chamber following intravitreal injection relative to the clearance of aflibercept from the ocular chamber following intravitreal injection of <4 mg aflibercept, wherein the method comprises intravitreally injecting into an eye of a subject in need thereof a single initial dose of about 8 mg or more of aflibercept, followed by

[0053] One or more second doses of aflibercept of about 8 mg or more, followed by

[0054] one or more third doses of aflibercept of about 8 mg or more;

[0055] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0056] wherein each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

[0057] The present invention provides aflibercept for use in a method for increasing the time at which the amount of free aflibercept in the ocular chamber of a subject reaches the lower limit of quantitation (LLOQ) following 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 following intravitreal injection of about 2 mg of aflibercept,

[0058] The method comprises intravitreally injecting a single initial dose of about 8 mg or more of aflibercept into the eye of a subject in need thereof, followed by

[0059] One or more second doses of aflibercept of about 8 mg or more, followed by

[0060] one or more third doses of aflibercept of about 8 mg or more;

[0061] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0062] wherein each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

[0063] The present invention provides aflibercept for use in 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 of aflibercept, wherein the method comprises 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 wherein each third dose is administered about 12-20 weeks after the immediately preceding dose.

[0064] The present invention provides a VEGF receptor fusion protein for use in the following method for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof,

[0065] - Improve the best corrected visual acuity of a subject in need with DR and / or DME; or

[0066] - Promote retinal dryness in subjects in need thereof with DR and / or DME;

[0067] Wherein the method comprises administering to the eye of the subject one or more doses of about 8 mg or more of a VEGF receptor fusion protein once every 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, or 20 weeks, or 12-20 weeks, or 12-16 weeks, or 16-20 weeks.

[0068] The present invention provides aflibercept for use in the treatment or prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the treatment or prevention comprises initiating treatment with a monthly (every 4 weeks) injection of 8 mg aflibercept 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 volume of application for 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 outcomes, such as, but not limited to, letter gain or letter loss in BCVA; increase or decrease in CRT; presence or absence of subretinal fluid; or presence or absence of hemorrhage or persistent or worsening DME. In one embodiment of the present invention, if the subject has been identified as a subject 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 or > 10 letters; CRT > 300 or 320 μm; CRT increase of 50 μm; or 2. persistent or worsening DME. In one embodiment of the present invention, if the subject has been identified as a subject 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, < 10 letters; CRT < 300 or 320 μm; CRT decrease > 50 μm; absence of subretinal fluid; or absence of hemorrhage.

[0069] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic macular edema in a subject in need thereof, wherein the method comprises administering 8 mg of 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 of VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 8-16 weeks (2-4 months + / - 7 days) or every 8-20 weeks (2-5 months + / - 7 days).

[0070] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy (DR) in a subject in need thereof, wherein the method comprises administering 8 mg of 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 of VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 8-16 weeks (2-4 months + / - 7 days) or every 8-20 weeks (2-5 months + / - 7 days).

[0071] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic macular edema (DME) in a subject in need thereof, wherein the method comprises administering 8 mg of the 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 of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 12 weeks (2-4 months + / - 7 days).

[0072] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy (DR) in a subject in need thereof, wherein the method comprises administering 8 mg of 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 of VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 12 weeks (2-4 months + / - 7 days).

[0073] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic macular edema (DME) in a subject in need thereof, wherein the method comprises administering 8 mg of the 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 of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 16 weeks (2-4 months + / - 7 days).

[0074] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy (DR) in a subject in need thereof, wherein the method comprises administering 8 mg of 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 of VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 16 weeks (2-4 months + / - 7 days).

[0075] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic macular edema (DME) in a subject in need thereof, wherein the method comprises administering 8 mg of the 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 of the VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 20 weeks (2-4 months + / - 7 days).

[0076] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy (DR) in a subject in need thereof, wherein the method comprises administering 8 mg of 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 of VEGF receptor fusion protein (0.07 mL) via intravitreal injection once every 20 weeks (2-4 months + / - 7 days).

[0077] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof:

[0078] (1) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein, and then the method comprises administering to the subject the initial 8 mg dose 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; and 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 a dosing schedule of HDq12, HDq16, or HDq20;

[0079] (2) wherein 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, 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;

[0080] (3) wherein the subject has received an initial 2 mg dose of VEGF receptor fusion protein, and the method comprises administering to the subject a second 8 mg dose of VEGF receptor fusion protein one month later, and 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 schedule;

[0081] (4) wherein the subject has received an initial 2 mg dose of the VEGF receptor fusion protein, and then the method comprises administering to the subject a first 8 mg maintenance dose of the VEGF receptor fusion protein one month later, and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing schedule;

[0082] (5) wherein 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 an initial 8 mg dose 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; and 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;

[0083] (6) wherein 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 first 8 mg second dose of VEGF receptor fusion protein one month later, and administering a second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0084] (7) wherein 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 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 schedule;

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

[0086] (9) wherein 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, then the method comprises administering to the subject an initial 8 mg dose 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; and 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;

[0087] (10) wherein 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, then the method comprises administering to the subject 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 thereafter; then, thereafter, 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;

[0088] (11) wherein 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, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein one month later, and 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;

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

[0090] (13) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein one month later and administering the first 8 mg second dose of VEGF receptor fusion protein one month thereafter; and administering the second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0091] (14) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject 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 thereafter; then, thereafter, 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;

[0092] (15) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein one month later; then, thereafter, 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;

[0093] (16) wherein 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 third 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 schedule;

[0094] (17) wherein 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 and a third 2 mg second dose of VEGF receptor fusion protein one month later; and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months later and a first 8 mg second dose of VEGF receptor fusion protein one month thereafter; and a second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0095] (18) wherein 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 and a third 2 mg second dose of VEGF receptor fusion protein one month later; and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein two months later and 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 thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0096] (19) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein two months later and thereafter administering a maintenance dose of 8 mg VEGF receptor fusion protein once or more for 12 weeks or 16 weeks according to the HDq12 or HDq16 or HDq20 dosing regimen for 12 weeks or 16 weeks thereafter;

[0097] (20) wherein 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 third 2 mg second dose of VEGF receptor fusion protein 1 month later and received a fourth 2 mg second dose of VEGF receptor fusion protein 1 month later; thereafter, the method then 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;

[0098] (21) wherein 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 and a third 2 mg second dose of VEGF receptor fusion protein one month later and a fourth 2 mg second dose of VEGF receptor fusion protein one month later; and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months after the last VEGF receptor fusion protein maintenance dose and, one month thereafter, administering the first 8 mg second dose of VEGF receptor fusion protein; and, one month thereafter, administering the second 8 mg second dose of VEGF receptor fusion protein; and 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;

[0099] (22) wherein 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 and a third 2 mg second dose of VEGF receptor fusion protein one month later and a fourth 2 mg second dose of VEGF receptor fusion protein one month later; and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein two months after the last VEGF receptor fusion protein maintenance dose and a second 8 mg second 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 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0100] (23) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein two months after the last maintenance dose of VEGF receptor fusion protein and thereafter administering one or more weekly 8 mg doses of VEGF receptor fusion protein for 12 weeks, 16 weeks or 20 weeks thereafter according to the HDq12, HDq16 or HDq20 dosing regimen;

[0101] or

[0102] (24) wherein 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 third 2 mg second dose of VEGF receptor fusion protein 1 month later and received a fourth 2 mg second dose of VEGF receptor fusion protein 1 month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein 2 months after the last VEGF receptor fusion protein maintenance dose 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;

[0103] in,

[0104] (i) The HDq12 dosing regimen includes:

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

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

[0107] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0108] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0109] wherein each third dose is administered approximately 12 weeks after the immediately preceding dose;

[0110] (ii) The HDq16 dosing regimen includes:

[0111] A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by

[0112] One or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by

[0113] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0114] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0115] wherein each third dose is administered approximately 16 weeks after the immediately preceding dose;

[0116] and

[0117] (iii) The HDq20 dosing regimen includes:

[0118] A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by

[0119] One or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by

[0120] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0121] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0122] wherein each third dose is administered approximately 20 weeks after the immediately preceding dose.

[0123] The present invention provides aflibercept for use in the treatment or prevention of diabetic macular edema or diabetic retinopathy in a subject in need thereof, the treatment or prevention comprising administering to the subject's eye a single initial dose of 8 mg of aflibercept followed by one or more third doses of about 8 mg of aflibercept; wherein each third dose is administered about 8, 12, 16, or 20 weeks after the immediately preceding dose. In one embodiment of the invention, the subject is not a treatment naive subject, or the subject has been previously treated with a VEGF antagonist, or preferably the subject has been previously treated with 8 mg of aflibercept or 2 mg of aflibercept.

[0124] The present invention provides aflibercept for use in the treatment or prevention of diabetic macular edema or diabetic retinopathy in a subject pretreated with 2 mg of aflibercept, the treatment or prevention comprising administering to the eye of the subject a single initial dose of about 8 mg of aflibercept, followed by one or more second doses of about 8 mg of aflibercept, followed by one or more third doses of about 8 mg of aflibercept, wherein each second dose is administered about 4 weeks after the immediately preceding dose and wherein each third dose is administered about 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 of aflibercept are administered to the eye of the subject according to a HDq12, HDq16, HDq20, or treatment and extended dosing schedule.

[0125] The present invention provides a VEGF receptor fusion protein for use in the treatment or prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the subject having received a dosing regimen for treating or preventing the condition, wherein:

[0126] (a) the subject has received an initial dose of 8 mg of a VEGF receptor fusion protein, and the method comprises administering to the subject a first 8 mg second dose of the VEGF receptor fusion protein one month later and a second 8 mg second dose of the VEGF receptor fusion protein one month thereafter; and then administering one or more 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0127] or

[0128] (b) the subject has received an initial dose of 8 mg of the VEGF receptor fusion protein and received a first 8 mg second dose of the VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a second 8 mg second dose of the VEGF receptor fusion protein one month later, and then administering one or more 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks thereafter according to the HDq12, HDq16, or HDq20 dosing regimen;

[0129] or

[0130] (c) the subject has received an initial 8 mg dose of a VEGF receptor fusion protein and received a first 8 mg second dose of the VEGF receptor fusion protein one month later and a second 8 mg second dose of the VEGF receptor fusion protein one month later, and then the method comprises administering to the subject a first 8 mg maintenance dose of the VEGF receptor fusion protein after 12 weeks, 16 weeks, or 20 weeks, and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing schedule;

[0131] or

[0132] (d) wherein the subject has received an initial 8 mg dose of a VEGF receptor fusion protein and one month later received a first 8 mg second dose of a VEGF receptor fusion protein and one month later received a second 8 mg second dose of a VEGF receptor fusion protein, and then every 12 weeks, 16 weeks, or 20 weeks thereafter, the subject has received one or more 8 mg maintenance doses of a VEGF receptor fusion protein; and then the method comprises administering to the subject one or more 8 mg maintenance doses of a VEGF receptor fusion protein 12 weeks, 16 weeks, or 20 weeks after the last maintenance dose of a VEGF receptor fusion protein and administering all other 8 mg maintenance doses of a VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen;

[0133] in,

[0134] (i) The HDq12 dosing regimen includes:

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

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

[0137] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0138] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0139] wherein each third dose is administered approximately 12 weeks after the immediately preceding dose;

[0140] (ii) The HDq16 dosing regimen includes:

[0141] A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by

[0142] One or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by

[0143] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0144] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0145] wherein each third dose is administered approximately 16 weeks after the immediately preceding dose;

[0146] and

[0147] (iii) The HDq20 dosing regimen includes:

[0148] A single initial dose of about 8 mg or more of VEGF receptor fusion protein, followed by

[0149] One or more second doses of about 8 mg or more of the VEGF receptor fusion protein, followed by

[0150] one or more third doses of about 8 mg or more of the VEGF receptor fusion protein;

[0151] wherein each second dose is administered about 2 to 4 weeks after the immediately preceding dose; and

[0152] wherein each third dose is administered approximately 20 weeks after the immediately preceding dose.

[0153] The present invention provides a VEGF receptor fusion protein for use in treating or preventing an angiogenic eye disease in a subject in need thereof, the subject having received a dosing regimen for treating or preventing the disease, the dosing regimen requiring: a single initial dose of about 2 mg of the 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 comprising administering to the subject's eye an 8 mg dose of the VEGF receptor fusion protein, evaluating the subject about 8 weeks, or 10 weeks, or 12 weeks after said administration, and continuing to administer 8 mg to the subject every 12 weeks or every 16 weeks if the treating physician determines that dosing every 12 weeks or every 16 weeks is appropriate. VEGF receptor fusion protein; or evaluating the subject within about 8 weeks, 10 weeks, or 12 weeks after said administration, and if the treating physician determines that dosing every 12 weeks is appropriate, administering another 8 mg dose of VEGF receptor fusion protein, re-evaluating the subject within about 12 weeks, and if the treating physician determines that dosing every 16 weeks is appropriate, continuing to administer 8 mg of VEGF receptor fusion protein to the subject every 16 weeks.

[0154] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy or diabetic macular edema in a subject in need thereof, wherein the treatment or prevention comprises administering to the subject's eye a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by one or more second doses (preferably 2 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 or 16 weeks after the immediately preceding dose; the method further comprises administering the third dose at intervals of about 12 or 16 weeks after the immediately preceding dose, after receiving one or more of the third doses about 12 or 16 weeks after the immediately preceding dose.

[0155] Extended from 12 weeks to 16 weeks;

[0156] · from 12 weeks to 20 weeks; or

[0157] Extended from 16 weeks to 20 weeks,

[0158] This is after the immediately preceding dose.

[0159] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the treatment or prevention comprising administering to the subject's eye 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, 16 weeks, or 20 weeks after the immediately preceding dose; the method further comprises administering the third dose at intervals of about 12 weeks, 16 weeks, or 20 weeks after the immediately preceding dose, after receiving one or more of the third doses.

[0160] Shortened from 12 weeks to 8 weeks;

[0161] From 16 weeks to 12 weeks;

[0162] From 16 weeks to 8 weeks,

[0163] From 20 weeks to 8 weeks,

[0164] from 20 weeks to 12 weeks, or

[0165] From 20 weeks to 16 weeks.

[0166] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the treatment and prevention comprising administering to the subject's eye three doses of about 8 mg of the VEGF receptor fusion protein in the form of a formulation comprising about 114.3 mg / ml of the VEGF receptor fusion protein at intervals of once every four weeks; wherein after the three doses, one or more doses of the VEGF receptor fusion protein are administered at intervals extending to 12 weeks, 16 weeks, or 20 weeks.

[0167] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the treatment and prevention comprising administering to the subject's eye a single initial dose of about 8 mg or more of the VEGF receptor fusion protein, followed by two 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 doses,

[0168] a) determining whether the subject meets at least one criterion for reducing or extending one or more intervals between doses of the VEGF receptor fusion protein by 2 weeks, 3 weeks, 4 weeks, or 2-4 weeks; and

[0169] b) if said determination is made, administering further doses of the VEGF receptor fusion protein at said reduced or extended dosage intervals,

[0170] The criteria for extending this interval include:

[0171] 1. BCVA loss < 5 letters; and / or

[0172] 2. CRT < 300 or 320 microns;

[0173] And, wherein the criteria for reducing the interval include:

[0174] 1. BCVA loss >10 letters;

[0175] 2. Persistent or worsening DME; and / or

[0176] 3. CRT increases by >50 microns.

[0177] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the subject having been pre-treated with one or more 2 mg doses of the VEGF receptor fusion protein, the treatment and prevention comprising administering to the subject's eye 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 12-20 weeks after the immediately preceding dose.

[0178] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of angiogenic eye diseases in a subject in need thereof, the treatment and prevention comprising administering to the eye of the subject

[0179] (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

[0180] (2) Administering a dose of 8 mg or more of VEGF receptor fusion protein once or more approximately every 4 weeks.

[0181] A VEGF receptor fusion protein for use in the treatment and prevention of angiogenic eye diseases, wherein the treatment or prevention comprises providing

[0182] One 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;

[0183] ·1 No. 18 1 / 2-inch, 5-micron filter needle, which includes a tip and bevel;

[0184] Pack together; then

[0185] (1) visually inspect the aqueous formulation in the vial and, if particulates, turbidity, or discoloration are visible, use another vial of the aqueous formulation containing the VEGF receptor fusion protein;

[0186] (2) removing the protective plastic cap from the vial; and

[0187] (3) Clean the top of the vial with an alcohol wipe; then

[0188] Use aseptic technique:

[0189] (4) Take out the No. 18 x 1 from their packaging 1 / 2 inch, 5 micron filter needle and the 1 mL syringe;

[0190] (5) attaching the filter needle to the syringe by screwing the filter needle onto the Luer lock syringe tip;

[0191] (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;

[0192] (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;

[0193] (8) Continue to tilt the vial during extraction to keep the bevel of the filter needle immersed in the formulation;

[0194] (9) pulling the plunger rod back sufficiently when emptying the vial to completely empty the filter needle;

[0195] (10) removing the filter needle from the syringe and disposing of the filter needle;

[0196] (11) removing the 30 gauge x 1 / 2 inch injection needle from its packaging and attaching the injection needle to the syringe by screwing it firmly onto the tip of the Luer lock syringe;

[0197] (12) holding the syringe with the needle pointing upward, and checking the syringe for bubbles, wherein if there are bubbles, tap the syringe gently with your fingers until the bubbles rise to the top; and

[0198] (13) Slowly depress the plunger until the plunger tip is aligned with the 70 μl mark on the syringe.

[0199] The present invention provides VEGF receptor fusion proteins for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the subject has been receiving a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema, the dosing regimen requiring: a single initial dose of about 2 mg of the VEGF receptor fusion protein, followed by four 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; wherein the subject is at any stage (initial dose, second dose, or third dose) of the 2 mg VEGF receptor fusion protein dosing regimen.

[0200] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 103-126 mg / ml of the VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0201] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising approximately 114.3 mg / ml VEGF receptor fusion protein, a histidine-based buffer, and arginine.

[0202] The present invention provides aflibercept for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein >8 mg of 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.

[0203] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein >8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising:

[0204] at least about 100 mg / ml of a VEGF receptor fusion protein;

[0205] About 10mM-100mM L-arginine;

[0206] sucrose;

[0207] Histidine-based buffers; and

[0208] surfactants;

[0209] wherein the pH of the formulation is from 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.

[0210] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising

[0211] >100 mg / ml VEGF receptor fusion protein, histidine-based buffer, and L-arginine;

[0212] 140 mg / ml aflibercept; 20 mM histidine-based buffer; 5% sucrose; 0.03% polysorbate 20; 10 mM L-arginine; pH 5.8;

[0213] 150 + 15 mg / ml aflibercept, 10 mM phosphate-based 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;

[0214] 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;

[0215] 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;

[0216] 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;

[0217] >100 mg / ml aflibercept, histidine-based buffer, and L-arginine;

[0218] >100 mg / ml aflibercept, approximately pH 5.8, wherein the formulation forms <3% HMW aggregates after incubation at 5°C for 2 months;

[0219] Approximately 114.3 mg / mL aflibercept; 10 mM-50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, pH 5.8;

[0220] 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

[0221] Approximately 114.3 mg / mL aflibercept; arginine monohydrochloride; histidine; histidine hydrochloride monohydrate; polysorbate 20; sucrose, and water for injection.

[0222] The present invention provides VEGF receptor fusion proteins for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the subject achieves and / or maintains one or more of the following:

[0223] Improvement in the Diabetic Retinopathy Severity Scale (DRSS), e.g., at least grade 2 or 3;

[0224] Improvement in best-corrected visual acuity;

[0225] Dry retina

[0226] Improvement in best-corrected visual acuity;

[0227] Improvement in best corrected visual acuity by at least 5, 10, or 15 letters

[0228] A BCVA of at least 69 letters;

[0229] Decreased central retinal thickness (CRT);

[0230] Absence of vascular leakage as measured by fluorescein angiography (FA);

[0231] Improvement in the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) total score compared to pre-treatment baseline;

[0232] The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea and in central subregions;

[0233] Maintaining a fluid-free retina (total fluid, IRF, and / or SRF in the foveal center and central subregion);

[0234] No macular edema;

[0235] Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT); and / or

[0236] Do not leave the HDq12 or HDq16 treatment regimen once started.

[0237] The present invention provides VEGF receptor fusion proteins for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the subject achieves and / or maintains one or more of the following:

[0238] Non-inferiority of BVCA compared with intravitreal aflibercept 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, followed by 2 mg approximately every 8 weeks or every 2 months;

[0239] An increase in BCVA (based on ETDRS letter score) of approximately 7, 8, or 9 letters from the start of treatment to week 60, with a baseline BCVA of approximately 61, 62, or 63;

[0240] BCVA (based on ETDRS letter scores) of at least about 69 letters from the start of treatment to Week 48 or Week 60;

[0241] No loss of 5, 10, 15, or 69 letters or more of BCVA after 12, 24, 36, 49, 60, 72, 84, or 90 weeks from the start of treatment;

[0242] Improvement in BCVA (based on ETDRS letter score) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0243] Improvement in BVCA from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48;

[0244] BCVA score (based on ETDRS letter score) of approximately 69, 70, 71, 72, or 73 between weeks 48 and 60;

[0245] A change in BCVA score (based on ETDRS letter score) of about 7, 8, or 9 from the start of treatment between Week 36 and Week 48, with a BCVA of about 60 or 70 at any point between Week 36 and Week 48;

[0246] A change in BCVA score (based on ETDRS letter score) from the start of treatment of about 7, 8, or 9 between Weeks 48 and 60, with a BCVA of about 69, 70, 71, 72, or 73 at any point between Weeks 48 and 60;

[0247] An increase in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity chart or Snellen equivalent of ≥4 letters, ≥5 letters, ≥6 letters, ≥7 letters, ≥8 letters, >9 letters, or >10 letters from the start of treatment to Weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48;

[0248] No loss of 5, 10, or 15 letters (based on the ETDRS letter score) from the start of treatment to week 48 or week 60;

[0249] An improvement of 5, 10, or 15 letters (based on the ETDRS letter score) from the start of treatment to week 48 or week 60;

[0250] BCVA improvement 4 weeks after treatment initiation of approximately 4 or 5 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 4 or 5 letters (ETDRS or Snellen equivalent) with HDq16;

[0251] BCVA improvement 8 weeks after treatment initiation of approximately 6 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0252] BCVA improvement 12 weeks after treatment initiation of approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0253] BCVA improvement 16 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq12, or 7 letters (ETDRS or Snellen equivalent) with HDq16;

[0254] Improvement in BCVA 20 weeks after treatment initiation of approximately 6 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0255] Improvement in BCVA 24 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0256] Improvement in BCVA 28 weeks after treatment initiation of approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0257] Improvement in BCVA 32 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0258] Improvement in BCVA 36 weeks after treatment initiation of 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16

[0259] Improvement in BCVA 40 weeks after treatment initiation was approximately 8 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16;

[0260] Improvement in BCVA 44 weeks after treatment initiation was approximately 8 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0261] Improvement in BCVA 48 weeks after treatment initiation of approximately 8 or 9 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0262] Improvement in BCVA approximately 8 weeks after the start of treatment and maintained thereafter until at least 48 weeks during the treatment regimen;

[0263] BCVA 4 weeks after the start of treatment is approximately 68 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0264] BCVA 8 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0265] BCVA 12 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0266] BCVA 16 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0267] BCVA 20 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0268] BCVA 24 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0269] BCVA 28 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0270] BCVA 32 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0271] BCVA 36 weeks after the start of treatment of approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0272] BCVA 40 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0273] BCVA 44 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0274] BCVA 48 weeks after the start of treatment is approximately 73 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0275] With HDq12, when baseline BCVA was approximately <73 ETDRS letters, the BCVA improvement at week 48 after treatment initiation was approximately 9 or 10 letters (ETDRS or Snellen equivalent);

[0276] When taking HDq12, when baseline BCVA is approximately >73 ETDRS letters, an improvement in BCVA of approximately 5 or 6 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0277] When taking HDq16, when baseline BCVA is approximately <73 ETDRS letters, an improvement in BCVA of approximately 8 or 9 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0278] When taking HDq16, when baseline BCVA is approximately >73 ETDRS letters, an improvement in BCVA of approximately 4 or 5 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0279] An improvement in BCVA of approximately 7 or 8 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is approximately < 400 microns on the HDq12 regimen;

[0280] When using the HDq12 regimen, when the baseline CRT is approximately >400 microns, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0281] An improvement in BCVA of approximately 5 or 6 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is approximately < 400 microns on the HDq16 regimen;

[0282] An improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is > approximately 400 microns on HDq16;

[0283] Increase in BCVA (based on ETDRS letter score) by >5, >10, or ≥15 letters from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0284] Improvement of ≥2 or >3 grades on the Diabetic Retinopathy Severity Scale (DRSS) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0285] Improvement of ≥ 2 grades on the Diabetic Retinopathy Severity Scale (DRSS) from the start of treatment to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, or Week 48;

[0286] Absence of retinal fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea and in central subregions as measured by optical coherence tomography (OCT) from the start of treatment to Weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0287] No vascular leakage as measured by fluorescein angiography (FA) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0288] Maintenance of retinal fluid-freeness (total fluid, IRF, and / or SRF in the foveal center and central subregion) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0289] A decrease in the total area (mm2) of fluorescein leakage within the ETDRS mesh by approximately 12, 13, or 14 mm2 or more at week 48 or week 60, as measured by fluorescein angiography;

[0290] Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0291] The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48;

[0292] Retinal dryness from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0293] Absence of central foveal fluid as measured by optical coherence tomography (OCT) from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0294] The change in central retinal thickness at 4 weeks after the start of treatment was approximately -118 or -118.3 microns on the HDq12 regimen; or approximately -124 or -125 or -124.9 or -125.5 microns on the HDq16 regimen;

[0295] The change in central retinal thickness at 8 weeks after the start of treatment was approximately -137 or -137.4 microns on the HDq12 regimen; or approximately -139 or -140 or -139.6 or -140.3 microns on the HDq16 regimen;

[0296] The change in central retinal thickness 12 weeks after the start of treatment was approximately -150 or -150.1 microns on the HDq12 regimen; or approximately -152 or -153 or -152.7 or -153.4 microns on the HDq16 regimen;

[0297] The change in central retinal thickness 16 weeks after the start of treatment was approximately -139 or -139.4 microns on the HDq12 regimen; or approximately -145 or -146 or -145.5 or -146.4 microns on the HDq16 regimen;

[0298] The change in central retinal thickness 20 weeks after the start of treatment was approximately -117 or -117.1 microns on the HDq12 regimen; or approximately -112 or -113 or -112.5 or -113.3 microns on the HDq16 regimen;

[0299] The change in central retinal thickness 24 weeks after the start of treatment was approximately -158 or -158.1 microns on the HDq12 regimen; or approximately -103 or -104 or -103.8 or -104.3 microns on the HDq16 regimen;

[0300] The change in central retinal thickness 28 weeks after the start of treatment was approximately -146, -147, or -146.7 microns on the HDq12 regimen; or approximately -162 or -162.3 microns on the HDq16 regimen;

[0301] The change in central retinal thickness 32 weeks after the start of treatment was approximately -132 microns on the HDq12 regimen; or approximately -145, -146, or -145.8 microns on the HDq16 regimen;

[0302] The change in central retinal thickness 36 weeks after the start of treatment was approximately -168 or -168.1 microns on the HDq12 regimen; or approximately -124 or -125 or -124.7 or -125.2 microns on the HDq16 regimen;

[0303] The change in central retinal thickness 40 weeks after the start of treatment was approximately -163 microns on the HDq12 regimen; or approximately -122, -123, -122.5, or -123.1 microns on the HDq16 regimen;

[0304] The change in central retinal thickness at 44 weeks after the start of treatment was approximately -147, -148, or -147.4 microns on the HDq12 regimen; or approximately -164, -164.1, or -164.3 microns on the HDq16 regimen;

[0305] The change in central retinal thickness at 48 weeks after the start of treatment was approximately -171, -172, or -171.7 microns on the HDq12 regimen; or approximately -148, -149, -148.3, or -149.4 microns on the HDq16 regimen;

[0306] The change in central retinal thickness 60 weeks after the start of treatment was approximately -181.95 or -176.24 microns on the HDq12 regimen; or approximately -166.26 or -167.18 microns on the HDq16 regimen;

[0307] The change in central retinal thickness between the start of treatment (week 0) and week 4 on the HDq12 regimen was approximately -118, -119, or -118.3 microns;

[0308] A change in central retinal thickness of approximately -19, -20, or -19.1 microns between weeks 4 and 8 on the HDq12 regimen;

[0309] A change in central retinal thickness of approximately -12, -13, or -12.7 microns between weeks 8 and 12 on the HDq12 regimen;

[0310] A change in central retinal thickness of approximately -40 or -41 microns between weeks 20 and 24 on the HDq12 regimen;

[0311] A change in central retinal thickness of approximately -36, -37, or -36.1 microns between weeks 32 and 36 on the HDq12 regimen;

[0312] A change in central retinal thickness of approximately -24, -25, or -24.3 microns between weeks 44 and 48 on the HDq12 regimen;

[0313] A change in central retinal thickness of -4, -5, or -4.5 microns between weeks 48 and 60 on the HDq12 regimen;

[0314] A change in central retinal thickness of approximately -124, -125, or -124.9 microns between the start of treatment (week 0) and week 4 on the HDq16 regimen;

[0315] A change in central retinal thickness of approximately -14, -15, or -14.7 microns between weeks 4 and 8 on the HDq16 regimen;

[0316] A change in central retinal thickness of approximately -13, -14, or -13.1 microns between weeks 8 and 12 on the HDq16 regimen;

[0317] A change in central retinal thickness of approximately -58, -59, or -58.5 microns between weeks 24 and 28 on the HDq16 regimen;

[0318] A change in central retinal thickness of approximately -41, -42, or -41.6 microns between weeks 40 and 44 on the HDq16 regimen;

[0319] A decrease in central retinal thickness at 4, 5, 6, 7, or 8 weeks after the initiation of treatment, which is then maintained within approximately +17, +18, or +19 microns during the treatment regimen and through at least 48 weeks after the initiation of treatment;

[0320] A decrease in central retinal thickness of approximately 100, 125, 150, 175, or 200 microns from the start of treatment to weeks 12, 24, 36, 49, 60, 72, 84, or 90;

[0321] a reduction in central retinal thickness of approximately 148 to 182 microns as measured by optical coherence tomography (OCT) at approximately week 48 or 60 after the start of treatment, where the baseline CRT is approximately 449, 450, 455, or 460 microns;

[0322] a decrease in central retinal thickness (CRT) of at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 microns from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48;

[0323] Free aflibercept in plasma was approximately 0.149 (±0.249) mg / L at approximately 0.1667 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0324] free aflibercept in plasma was approximately 0.205 (±0.250) mg / l at approximately 0.3333 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0325] About 0.266 (±0.211) mg / L free aflibercept in plasma at about 1 day after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0326] About 0.218 (±0.145) mg / L free aflibercept in plasma at about 2 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0327] About 0.140 (±0.0741) mg / L free aflibercept in plasma about 4 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0328] approximately 0.0767 (±0.0436) mg / L free aflibercept in plasma approximately 7 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0329] About 0.0309 (±0.0241) mg / l free aflibercept in plasma at about 14 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0330] approximately 0.0171 (±0.0171) mg / L free aflibercept in plasma at approximately 21 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0331] approximately 0.00730 (±0.0113) mg / L free aflibercept in plasma at approximately 28 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks;

[0332] At approximately 0.1667 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.00698 (±0.0276) mg / L; whereas at baseline, there was approximately 0.00583 mg / L (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0333] At approximately 0.3333 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.00731 (±0.0279) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0334] About 0.0678 (±0.0486) mg / L adjusted bound aflibercept in plasma about 1 day after the first dose; wherein at baseline, there was about 0.00583 mg / L (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0335] Approximately 2 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.138 (±0.0618) mg / l; at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0336] Approximately 4 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.259 (±0.126) mg / l; at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks;

[0337] Approximately 7 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.346 (±0.151) mg / l; at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subjects had not received intravitreal aflibercept treatment for at least 12 weeks;

[0338] Approximately 14 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.374 (±0.110) mg / l; at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subjects had not received intravitreal aflibercept treatment for at least 12 weeks;

[0339] Approximately 21 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.343 (±0.128) mg / l; at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subjects had not received intravitreal aflibercept treatment for at least 12 weeks;

[0340] About 28 days after the first dose, the adjusted bound aflibercept in plasma was about 0.269 (±0.149) mg / l; at baseline, there was about 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subjects had not received intravitreal aflibercept treatment for at least 12 weeks;

[0341] The maximum level of free aflibercept in plasma was reached approximately 0.965 days after the first dose.

[0342] Free aflibercept in the plasma reached a maximum level of approximately 0.310 mg / l (±0.263);

[0343] The free aflibercept in the plasma is from about 0 to about 1.08 mg / L;

[0344] The maximum free aflibercept level (mg / l) in the plasma per dose (mg) of aflibercept was approximately 0.388 (±0.0328) mg / l / mg;

[0345] The adjusted conjugated aflibercept level in plasma reached its maximum approximately 14 days after the first dose;

[0346] The adjusted bound aflibercept reached a maximum level in plasma of approximately 0.387 mg / l (±0.135);

[0347] The adjusted bound aflibercept in the plasma is about 0.137 to about 0.774 mg / L;

[0348] The adjusted maximum amount of bound aflibercept in the plasma (mg / l) per dose (mg) of aflibercept was approximately 0.483 (±0.0168) mg / l / mg;

[0349] No anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment;

[0350] Improvement in the National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score compared to pre-treatment baseline; and / or

[0351] No macular edema.

[0352] The present invention provides a VEGF receptor fusion protein for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein

[0353] Administering one initial dose, two secondary doses, and three tertiary doses of >8 mg of VEGF receptor fusion protein to the subject during the first year;

[0354] wherein the subject is administered one initial dose, two secondary doses, and two tertiary doses of the >8 mg VEGF receptor fusion protein during the first year; or

[0355] • wherein said subject is administered one initial dose, two secondary doses, and three tertiary doses of said >8 mg VEGF receptor fusion protein in the first year, followed by 2-4 tertiary doses in the second year.

[0356] The present invention provides VEGF receptor fusion proteins for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the dosage interval of 8 mg VEGF receptor fusion protein is adjusted (increased / maintained / decreased) based on visual and / or anatomical results.

[0357] The present invention provides VEGF receptor fusion proteins for use in the treatment and prevention of diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, wherein the doses of 8 mg of VEGF receptor fusion protein are administered according to a As Needed (PRN), Capped PRN, or Treat and Extended (T&E) dosing schedule.

[0358] The present invention also provides a kit comprising i) a container comprising a VEGF receptor fusion protein (preferably aflibercept) and ii) instructions for use of the VEGF fusion protein. In one embodiment of the present invention, the container is a vial or a prefilled syringe, wherein the vial is a Type I glass vial containing a nominal fill volume of approximately 0.26 mL of intravitreal injection solution. In one embodiment of the present invention, the container comprises a VEGF receptor fusion protein at a concentration greater than or equal to 100 mg / mL, or the container comprises aflibercept at a concentration of approximately 114.3 mg / mL. In one embodiment of the present invention, the instructions for use include instructions for using the VEGF fusion protein or aflibercept to treat DME and / or AMD. In one embodiment of the present invention, the instructions for use include 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 an amount to deliver a single dose of 70 microliters containing 8 mg of aflibercept to an adult patient, iii) the recommended dose is 8 mg of aflibercept (equivalent to 70 microliters of injection solution), iv) treatment with 8 mg of aflibercept is initiated with a monthly injection (every 4 weeks) for 3 consecutive doses; v) the injection interval can then be expanded to every 16 weeks or 20 weeks, vi) the treatment interval can be adjusted based on the physician's visual and / or anatomical judgment; and / or vii) 8 mg of aflibercept / 0.07 mL is provided as a sterile aqueous solution containing arginine monohydrochloride, histidine, histidine hydrochloride monohydrate, polysorbate 20, sucrose, and water for injection. BRIEF DESCRIPTION OF THE DRAWINGS

[0359] Figure 1 : Summary of PHOTON clinical trials.

[0360] Figure 2 Key eligibility criteria (inclusion and exclusion criteria) for the PHOTON clinical trial.

[0361] Figure 3 Dosing schedule and dose regimen modification (DRM) criteria for the PHOTON clinical trial.

[0362] Figure 4 Dose regimen modification criteria for the PHOTON clinical trial.

[0363] Figure 5 Patient disposition at week 48 of the PHOTON clinical trial.

[0364] Figure 6 Baseline demographics of subjects in the PHOTON clinical trial.

[0365] Figure 7 Baseline characteristics of the study eyes of participants in the PHOTON clinical trial.

[0366] Figure 8 Average number of injections through week 48 in the PHOTON clinical trial.

[0367] Figure 9 *Mean change in best-corrected visual acuity (BCVA) through Week 48 in the PHOTON clinical trial.

[0368] Shown are the least squares mean changes from baseline at Week 48.

[0369]

[0370] *The present invention includes methods for achieving approximately a specified improvement in BVCA by a specified time point while receiving a specified treatment regimen for treating DR and / or DME.

[0371] Figure 10 Percentage of subjects maintaining Q12-week and Q16 intervals through Week 48 in the PHOTON clinical trial.

[0372] Figure 11 The key secondary endpoint of the PHOTON clinical trial was the percentage of subjects achieving a ≥ 2-grade improvement on the Diabetic Retinopathy Severity Scale (DRSS) at Week 48.

[0373] Figure 12 Percentage of subjects with no retinal fluid in the center of the fovea at Week 48 in the PHOTON clinical trial.

[0374] Figure 13 *Mean change from baseline in central retinal thickness in the PHOTON clinical trial through Week 48. Various matched intervals are highlighted in the three insets.

[0375]

[0376] *The present invention includes methods for achieving approximately a specified reduction in central retinal thickness by a specified time point when receiving a specified treatment regimen for treating DR and / or DME.

[0377] Figure 14A 、 Figure 14B and Figure 14C : Serious treatment-emergent adverse events (TEAEs) of the eye in the PHOTON clinical trial up to Week 48 ( Figure 14A ); the most common adverse events (AEs) by week 48 ( Figure 14B ); non-ocular safety through week 48 ( Figure 14C ).

[0378] Figure 15 Intraocular inflammation during the PHOTON clinical trial through week 48 of treatment.

[0379] Figure 16 Mean change from baseline in intraocular pressure at week 48 in the PHOTON clinical trial.

[0380] Figure 17 Percentage of subjects meeting intraocular pressure criteria in the PHOTON clinical trial.

[0381] Figure 18 In the PHOTON clinical trial, non-ocular serious TEAEs were ≥ 1% through Week 48.

[0382] Figure 19 Antiplatelet Trialists' Collaboration (APTC) events occurred during the treatment period up to Week 48 of the PHOTON clinical trial.

[0383] Figure 20 Hypertensive events that occurred during the treatment period up to Week 48 in the PHOTON clinical trial.

[0384] Figure 21 Potentially clinically significant values ​​(PCSV) in blood pressure at week 48 in the PHOTON clinical trial.

[0385] Figure 22 Mean change from baseline in systolic blood pressure in the PHOTON clinical trial through Week 48. Baseline to Week 9 and mean baseline are shown in the inset.

[0386] Figure 23 Mean change from baseline in diastolic blood pressure in the PHOTON clinical trial through Week 48. Baseline to Week 9 and mean baseline are shown in the inset.

[0387] Figure 24 Death at week 48 in the PHOTON clinical trial.

[0388] Figure 25A and 25B :(A) Mean change from baseline in BCVA score (ETDRS letters) in study eyes at week 60, OC (Full analysis set); (B) Least squares mean difference in BCVA score (ETDRS letters) compared with baseline in the study eyes at week 60 (full analysis set). Value changes. Abbreviations: 2q8: 2 mg aflibercept administered every 8 weeks after 5 initial injections at 4-week intervals; HDq12: high-dose aflibercept 8 mg administered every 12 weeks after 3 initial injections at 4-week intervals; HDq16: high-dose aflibercept 8 mg administered every 16 weeks after 3 initial injections at 4-week intervals. BCVA = best-corrected visual acuity; ETDRS = Early Treatment Diabetic Retinopathy Study; HD = high dose; OC = observed case, SE = standard error. OC: observed after excluding the ICE defined for the primary estimator.

[0389] Figure 26A and 26B :(A) Mean change from baseline in central retinal thickness (micrometers) at the 60th week visit, OC (Full analysis set); (B) Least squares mean change from baseline in central retinal thickness (μm) at visit 60 weeks, OC (Full analysis set). Abbreviations: 2q8 = 2 mg aflibercept every 8 weeks after 5 initial injections at 4-week intervals; HDq12 = high-dose 8 mg aflibercept every 12 weeks after 3 initial injections at 4-week intervals; HDq16 = high-dose 8 mg aflibercept every 16 weeks after 3 initial injections at 4-week intervals. ICE = intermittent event; OC = observed case; SE = standard error. OC = observation after excluding the ICE defined for the primary estimator.

[0390] Figure 27 In an intensive PK sampling substudy (Study VGFTe-HD-DME-1934, logarithmic scale, [DPKS]), Mean (± 0.05) plasma free aflibercept levels by nominal time and treatment in unilaterally treated participants with DME SD) concentration (+SD) (mg / l). n = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = 5 initial monthly doses followed by intravitreal injections of 2 mg aflibercept every 8 weeks; HDq12 = 3 initial monthly doses followed by intravitreal injections of high-dose (8 mg) aflibercept every 12 weeks; HDq16 = 3 initial monthly doses followed by intravitreal injections of high-dose (8 mg) aflibercept every 16 weeks; LLOQ = lower limit of quantification; D = day; H = hour Note: The table below the figure presents the number of subjects contributing to the statistics for the corresponding visit / time point and treatment. Concentrations below the LLOQ were set as LLOQ / 2. HDq12 + HDq16 = combined data from treatment groups HDq12 and HDq16. Patients in the intensive PK substudy received only a single aflibercept injection.

[0391] Figure 28 In an intensive PK sampling substudy (Study VGFTe-HD-DME-1934, logarithmic scale, [DPKS]), Adjusted Bound Aflibercept in Plasma by Nominal Time and Treatment in Unilaterally Treated Participants with DME Mean (±SD) concentration (+SD) (mg / l).

[0392] n = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = 5 initial monthly doses followed by intravitreal injections of 2 mg aflibercept every 8 weeks; HDq12 = 3 initial monthly doses followed by intravitreal injections of high-dose (8 mg) aflibercept every 12 weeks; HDq16 = 3 initial monthly doses followed by intravitreal injections of high-dose (8 mg) aflibercept every 16 weeks; LLOQ = lower limit of quantification; D = day; H = hour. Note The table below the figure presents the number of subjects contributing to the statistics for the corresponding visit / time point and treatment. Concentrations below the LLOQ were set as LLOQ / 2. Adjusted bound aflibercept = 0.717*bound aflibercept. HDq12 + HDq16 = combined data from treatment groups HDq12 and HDq16. Patients in the intensive PK substudy received only a single aflibercept injection.

[0393] Figure 29 In a sparse PK sampling study (Study VGFTe-HD-DME-1934, logarithmic scale, [PKAS]), patients with DME Mean (±SD) concentrations of free aflibercept in plasma (mg / l)。 n = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = intravitreal injections of 2 mg aflibercept every 8 weeks after 5 initial monthly doses; HDq12 = intravitreal injections of high-dose (8 mg) aflibercept every 12 weeks after 3 initial monthly doses; HDq16 = intravitreal injections of high-dose (8 mg) aflibercept every 16 weeks after 3 initial monthly doses; LLOQ = lower limit of quantification; PKAS = pharmacokinetic analysis set. Note : The table below the figure presents the number of subjects who contributed to the statistics for the corresponding visit / time point and treatment group. Concentrations below the LLOQ were set as LLOQ / 2. Post-dose samples and samples collected during the intensive PK substudy (from day 0 to day 21 after dosing) were excluded.

[0394] Figure 30 In a sparse PK sampling study (Study VGFTe-HD-DME-1934, logarithmic scale, [PKAS]), patients with DME Adjusted mean (±SD) concentrations of conjugated aflibercept in plasma among participants by nominal time and treatment group (mg / l); N = number of subjects; SD = standard deviation; DME = diabetic macular edema; 2q8 = 5 intravitreal injections of 2 mg aflibercept every 8 weeks after the initial monthly dose; HDq12 = 3 intravitreal injections of high-dose (8 mg) aflibercept every 12 weeks after the initial monthly dose; HDq16 = 3 intravitreal injections of high-dose (8 mg) aflibercept every 16 weeks after the initial monthly dose; LLOQ = 3 / 4 lower limit of quantitation. Note: The table below the figure presents the number of subjects contributing to the statistics for the corresponding visit / time point and treatment group. Concentrations below the LLOQ were set as LLOQ / 2. Adjusted bound aflibercept = 0.717*bound aflibercept. Postdose samples and samples collected during the intensive PK substudy (from Day 0 to Day 21 after dosing) were excluded.

[0395] Figure 31 Structural representation of the population pharmacokinetic model for aflibercept following IV, SC, and IVT administration. 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 the subcutaneous depot compartment, K70 = elimination rate constant from the tissue (platelet) compartment; QE = intercompartmental clearance of free aflibercept between the ocular and central compartments, QF1 and QF2 = intercompartmental clearance of free aflibercept, VMK24, KM = saturable Michaelis-Menten type binding of free aflibercept to VEGF; VMK27, KMK27 = saturated elimination from the plasma compartment to the tissue compartment (platelets), CMT 2 and CMT 4 are both representatives of the plasma compartment and equal volumes are assumed.

[0396] Figure 32 In an intensive PK substudy (DPKS, logarithmic scale), single 2 mg and 8 mg IVT doses in nAMD or DME were compared with placebo. Mean (±SD) free and adjusted bound aflibercept concentrations (mg / l) over 28 days. LQ = below limit of quantitation, DME = diabetic macular edema, DPKS = intensive pharmacokinetic substudy, HDq12 = 8 mg aflibercept administered every 12 weeks after an initial three monthly injections, HDq16 = 8 mg aflibercept administered every 16 weeks after an initial three monthly injections, IVT = intravitreal, LLOQ = lower limit of quantitation, N = number of subjects, nAMD = neovascular age-related macular degeneration, SD = standard deviation. Adjusted bound aflibercept = 0.717*bound aflibercept Note: Concentrations below the LLOQ (free aflibercept concentration of 0.0156 mg / L, adjusted bound aflibercept concentration of 0.0224 mg / L) were set as LLOQ / 2. Note: Data for the first 28 days of 8 mg HD aflibercept (obtained from PULSAR or PHOTON) were a combination of data from participants who received HDq12 or HDq16. One participant in PULSAR who had an outlier free aflibercept concentration greater than 10 times the mean concentration at day 28 was excluded. Records from the ipsilateral eye after treatment were excluded. Data sources: Drug concentration data from the 48-week database locks for PULSAR and PHOTON and the final lock for CANDELA.

[0397] Figure 33 In an intensive PK substudy (DPKS), subjects with nAMD or DME were enrolled, stratified by dose and population. Observed and model concentrations of free and adjusted bound aflibercept in plasma over 28 days after a single IVT injection Type predicted concentration (mg / l). DME = diabetic macular edema, IVT = intravitreal, LLOQ = lower limit of quantification, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetic observation concentration below the lower limit of quantification (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) set at LLOQ / 2. Data source: Drug concentration data from intensive PK substudies in PHOTON, PULSAR, and CANDELA.

[0398] Figure 34 Observations of free and adjusted bound aflibercept in plasma in the combined nAMD and DME population Superposition of concentrations and model-predicted concentrations (mg / l).2q8 = 2 mg aflibercept every 8 weeks after 3 initial injections 4 weeks apart; 2q12 = 2 mg aflibercept every 12 weeks after 3 initial injections 4 weeks apart; 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 quantification, nAMD = neovascular age-related macular degeneration; observed concentrations below the lower limit of quantification (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Data source: Drug concentration data from CANDELA, PHOTON, and PULSAR.

[0399] Figure 35 In participants with combined nAMD and DME, stratified by dosing regimen, the model predicted a positive association after a single I·T injection. Measured aflibercept exposure (mg). DME = diabetic macular edema, HD = aflibercept 8 mg, IVT = intravitreal, nAMD = neovascular age-related macular degeneration, PI = prediction interval, PK = pharmacokinetics, QE = intercompartmental clearance of free aflibercept between the ocular and central compartments, with an adjusted LLOQ (0.0624 μg) set as the LLOQ of free aflibercept in plasma (i.e., 0.0156 mg / L) multiplied by the assumed volume of the study ocular compartment in the PK model (i.e., 4 mL). Because intraocular (free or bound) aflibercept concentrations were not measured in the clinical studies included in the population PK analysis dataset, this target was arbitrarily selected based on the LLOQ in plasma and used as a reference for comparisons between dosing regimens to assess the impact of HD aflibercept on QE.

[0400] Figure 36 In an intensive PK substudy (DPKS, logarithmic scale), single 2 mg and 8 mg IVT doses of dapoxetine were administered in participants with nAMD. Mean (±SD) free and adjusted bound aflibercept concentrations (mg / l) over 28 days for each dose of aflibercept Group value. DME = diabetic macular edema, DPKS = intensive pharmacokinetic substudy, HDq12 = 8 mg aflibercept administered every 12 weeks after an initial three monthly injections, HDq16 = 8 mg aflibercept administered every 16 weeks after an initial three monthly injections, IVT = intravitreal, LLOQ = lower limit of quantification, N = number of subjects, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetics, SD = standard deviation adjusted for combined aflibercept = 0.717* combined aflibercept. Note : The concentration below the LLOQ (free aflibercept concentration of 0.0156 mg / L and adjusted bound aflibercept concentration of 0.0224 mg / L) was set as LLOQ / 2. NoteData for the first 28 days of 8 mg (obtained from PULSAR or PHOTON) were combined from participants who received HDq12 or HDq16. Data sources: Drug concentrations at week 48 from PULSAR and PHOTON and at the final lock from CANDELA. Recordings after treatment of the ipsilateral eye were excluded.

[0401] Figure 37 In an intensive PK substudy (DPKS, logarithmic scale), single 2 mg and 8 mg IVT doses of dapoxetine were administered in participants with nAMD. Mean (±SD) free and adjusted bound aflibercept concentrations (mg / l) over 28 days for doses of aflibercept - including Outliers. DME = diabetic macular edema, DPKS = intensive pharmacokinetic substudy, HDq12 = 8 mg aflibercept administered every 12 weeks after an initial three monthly injections, HDq16 = 8 mg aflibercept administered every 16 weeks after an initial three monthly injections, IVT = intravitreal, LLOQ = lower limit of quantification, N = number of subjects, nAMD = neovascular age-related macular degeneration, PK = pharmacokinetics, SD = standard deviation adjusted for combined aflibercept = 0.717* combined aflibercept. Note Concentrations below the lower limit of quantification (LLOQ, free aflibercept concentration 0.0156 mg / L, adjusted bound aflibercept concentration 0.0224 mg / L) were set at LLOQ / 2. Data source: Drug concentrations at week 48 of PULSAR and the final lock of CANDELA. Data from VGFTOD-0702 were used as a reference (concentrations in the PK substudy >LLOQ were subtracted from predose concentrations). Recordings after treatment of the ipsilateral eye were excluded.

[0402] Figure 38 In an intensive PK substudy (DPKS, logarithmic scale), single 2 mg and 8 mg IVT doses were administered in participants with DME. Mean (±SD) free and adjusted bound aflibercept concentrations (mg / l) over 28 days for each dose of aflibercept. BLQ = below limit of quantification, DME = diabetic macular edema, DPKS = dense pharmacokinetic analysis set, HDq12 = 8 mg aflibercept administered every 12 weeks after an initial three monthly injections, HDq16 = 8 mg aflibercept administered every 16 weeks after an initial three monthly injections, IVT = intravitreal, LLOQ = lower limit of quantification, N = number of subjects, nAMD = neovascular age-related macular degeneration, SD = standard deviation Note : The concentration below LLOQ (free aflibercept concentration of 0.0156 mg / L, adjusted bound aflibercept concentration of 0.0224 mg / L) was set as LLOQ / 2. Adjusted bound aflibercept = 0.717*bound aflibercept. Note : Data for the first 28 days of 8 mg (obtained from PULSAR or PHOTON) were combined from data from participants who received HDq12 or HDq16. NoteIf participants took aflibercept within 12 weeks before starting study drug and their baseline concentration was > BLQ, this concentration was subtracted from the baseline concentration. Data source: Drug concentration data from the PHOTON Week 48 lock. Drug concentration data from VGFT-OD-0706 (historical data) were included as a reference. Recordings from the ipsilateral eye after treatment were excluded.

[0403] Figure 39 Observations of free and adjusted bound aflibercept in plasma of participants with nAMD Superposition of concentration and model-predicted concentration (mg / l) 2q8 = 2 mg aflibercept every 8 weeks after 3 initial injections 4 weeks apart; 2q12 = 2 mg aflibercept every 12 weeks after 3 initial injections 4 weeks apart; 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 quantification, nAMD = neovascular age-related macular degeneration; PK = pharmacokinetics, observed concentrations below the lower limit of quantification (LLOQ; 0.0156 mg / L for free aflibercept and 0.0224 mg / L for adjusted bound aflibercept) were set as LLOQ / 2. Data from PULSAR and CANDELA.

[0404] Figure 40 Free aflibercept and cytotoxicity in plasma of participants with diabetic macular edema in the PHOTON study Adjusted superposition of observed and model-predicted aflibercept concentrations (mg / l) 2q8 = 2 mg aflibercept every 8 weeks after 3 initial injections 4 weeks apart; HDq12 = 8 mg aflibercept every 12 weeks after 3 initial injections 4 weeks apart; HDq16 = 8 mg aflibercept every 16 weeks after 3 initial injections 4 weeks apart; IVT = intravitreal; LLOQ = lower limit of quantification. Data from PHOTON. DETAILED DESCRIPTION

[0405] The present invention provides, in part, a safe and effective high-dose IVT injection that extends the maintenance dosing interval to beyond 8 weeks, with at least similar functional and potentially improved anatomical outcomes. This regimen demonstrated unexpectedly high levels of durability in subjects, exceeding the expected durability based solely on the administration of increased aflibercept.

[0406] EYLEA has become the standard of care for diabetic macular edema (DME) and diabetic retinopathy (DR). Eylea is indicated for DME and DR at a dose of 2 mg once monthly for five doses, followed by maintenance dosing every eight weeks. The dosing regimens of the present invention have demonstrated that a significant percentage of subjects can maintain 12- 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 the consistent safety profile of EYLEA, support high-dose aflibercept as a potential new standard of care for angiogenic eye diseases such as DR or DME. The data presented herein demonstrate that 12- and 16-week dosing regimens of 8 mg aflibercept achieved high-barrier, sustained improvements in visual acuity and anatomical measures of retinal fluid over 48 weeks in subjects with diabetic macular edema. These results were achieved in subjects who promptly initiated the extended dosing interval, the vast majority of whom did not require regimen modification. In summary, the pivotal data support that aflibercept 8 mg provides a longer duration of action while maintaining a similar safety profile to EYLEA.

[0407] Prior to the initiation of the HD aflibercept clinical development program, pharmacokinetic simulations of the concentration-time profile of free aflibercept in the human vitreous using a single-chamber eye model predicted that an 8mg IVT dose of aflibercept would extend the dosing interval by approximately 20 days (two half-lives) relative to a 2mg IVT dose. The HDq12 and HDq16 regimens of aflibercept demonstrated a longer-lasting efficacy than predicted in HD clinical studies. Subsequent population PK analyses integrating data from the Phase 3 CANDELA PHOTON and PULSAR studies indicated that the intraocular clearance of free aflibercept with the HD drug product was 34% slower than that with 2mg IVT aflibercept administered as Eylea. This slower intraocular clearance of HD aflibercept is expected to result in longer intraocular persistence of free aflibercept and a duration of efficacy that is approximately 6 weeks longer than with the 2mg dose. The greater-than-expected reduction in ocular clearance of HD aflibercept compared to 2 mg Eylea was attributed to an "HD aflibercept effect," a highly statistically significant effect in a population PK model that could not be explained solely by increasing the dose from 2 mg to 8 mg.

[0408] Population PK predicted that the median time to reach the lower limit of quantification (LLOQ) for free aflibercept concentrations in plasma following 2 mg IV aflibercept was estimated to be 1.5 weeks, compared to 3.50 weeks for 8 mg HD aflibercept. The longer duration of systemic exposure to free aflibercept (representing movement of free aflibercept from the eye) with the HD aflibercept regimen was attributed not only to the higher administered dose and nonlinear systemic target-mediated elimination, but also to a 34% slower ocular clearance of free aflibercept. The slower ocular clearance of the HD aflibercept product was predicted to provide a 6-week longer duration of efficacy compared to the 2 mg aflibercept product, as the population PK estimated time to reach the amount of free aflibercept in the ocular compartment at the end of the 8-week dosing interval with the 2q8 regimen occurred after 6 weeks for the HD aflibercept product. Exposure-response analysis estimated that the slower ocular clearance of 8 mg aflibercept was attributable to the effects of the HD drug, resulting in a 20.6% lower dosing regimen modification (DRM) rate than would be expected if the HD drug had the same ocular clearance as 2 mg aflibercept.

[0409] Standard molecular biology methods are described by Sambrook, Fritsch and Maniatis (2nd ed. 1982 and 1989, 3rd ed. 2001) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; and Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, Calif. Standard methods also appear in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. New York, NY, which describes cloning in bacterial cells and DNA mutagenesis (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3), and bioinformatics (Vol. 4).

[0410] 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). Chemical analysis, chemical modifications, post-translational modifications, fusion protein production, protein glycosylation are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley & Sons, New York; Ausubel, et al. (2001) Current Techniques in Molecular Biology, Vol. 3, John Wiley & Sons, New York, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, Mo.; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, NJ, pp. 384-391). The production, purification, and fragmentation of polyclonal and monoclonal antibodies are described (Coligan et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley & Sons, New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard techniques for characterizing ligand / receptor interactions are available (see, e.g., Coligan et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley & Sons, New York).

[0411] Flow cytometric methods, including fluorescence-activated cell sorting (FACS), are available (see, e.g., Owens, et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley & Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, 2nd ed., Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley & Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies for use as, for example, diagnostic reagents are available (Molecular Probes (2003) catalog, Molecular Probes, Inc., Eugene, Oreg.; Sigma-Aldrich (2003) catalog, St. Louis, MO).

[0412] Standard immune system histological methods are described (see, e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams & Wilkins, Philadelphia, Pa.; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

[0413] "Isolated" VEGF antagonists and VEGF receptor fusion proteins (e.g., aflibercept), polypeptides, polynucleotides, and vectors 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 medium. An isolated VEGF antagonist or VEGF receptor fusion protein may further be at least partially free of expression system components, such as biomolecules from the host cells or their growth medium. In general, the term "isolated" is not intended to refer to the complete absence of such biomolecules (e.g., small or insignificant amounts of impurities may remain), or to the absence of water, buffers, or salts, or to components of a pharmaceutical formulation that includes a VEGF antagonist or VEGF receptor fusion protein.

[0414] 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. A subject or subject may be said to "suffer from" an angiogenic eye disease, such as nAMD. One or both eyes of such a subject or patient suffer from the disease. 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):

[0415] 1. ≥50 years old, such as 61, 62, 63, 74, or 75 years old;

[0416] 2. Active subfoveal CNV secondary to nAMD, including juxtafoveal lesions affecting the fovea of ​​the eye;

[0417] 3. Early Treatment Study for Diabetic Retinopathy with Best Corrected Visual Acuity (BCVA)

[0418] an (ETDRS) letter score of about 78 to 24, 73-78, <73, 58, 59, 60, 61, 62, or 63 (Snellen equivalent of 20 / 40, 20 / 63, 20 / 50, 20 / 32, or 20 / 320), e.g., due to DME or wet AMD;

[0419] 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 DME with central involvement in the eye, CRT ≥300 μm (or ≥320 μm as determined by Spectralis)

[0420] 5. The total lesion area is about 6 or 7 mm 2 , such as where the lesion type is occult, predominantly classic, or minimally classic;

[0421] 6. DRSS score better than or equal to 43, 47 or worse;

[0422] 7. Type 1 or type 2 diabetes mellitus (insulin-dependent or non-insulin-dependent) (e.g., about 15 years or longer);

[0423] 8. Hemoglobin A1C (%) of about 7 or 8 or higher;

[0424] 9. A body mass index of about 30 or 31 or higher; and / or

[0425] 10. History of diabetic retinedema, diabetic retinopathy, dry eye, vitreous detachment, retinopathy allergy, retinal hemorrhage, cataract surgery, retinal laser photocoagulation and intraocular lens implantation, hypertension, and / or the presence or absence of any one or more of the following characteristics:

[0426] 1. Signs of macular edema in the eye from any cause other than diabetes;

[0427] 2. IOP in the eye ≥ 25 mmHg;

[0428] 3. The eye has a history of glaucoma filtration surgery in the past or may need filtration surgery in the future;

[0429] 4. Any signs of infectious blepharitis, keratitis, scleritis or conjunctivitis in either eye within 4 weeks (28 days) of treatment

[0430] 5. Any intraocular inflammation and / or eye infection within 12 weeks (84 days) of treatment;

[0431] 6. History of idiopathic or autoimmune uveitis in the eyes;

[0432] 7. Vitreomacular traction or epiretinal membrane in the eye, such as that clearly visible on biomicroscopy or OCT, is thought to affect central vision;

[0433] 8. Preretinal fibrosis involving the macula in the eye;

[0434] 9. A history of macular hole of stage 2 or above;

[0435] 10. Current iris neovascularization, vitreous hemorrhage, or tractional retinal detachment as seen in the screening evaluation;

[0436] 11. The patient has a history of corneal transplantation or corneal dystrophy.

[0437] 12. Any concurrent ocular condition that, in the opinion of the treating physician, may increase the risk to the subject beyond that anticipated from the standard IVT injection procedure, or may otherwise interfere with the VEGF antagonist injection procedure;

[0438] 13. VEGF antagonists are contraindicated if there is reasonable suspicion of a disease or condition due to other medical history, metabolic dysfunction, physical examination results, or clinical laboratory findings;

[0439] 14. Any previous systemic (IV) anti-VEGF administration.

[0440] 15. Uncontrolled diabetes defined by hemoglobin A1c (HbA1c) > 12%;

[0441] 16. Uncontrolled blood pressure (defined as systolic blood pressure >160 mmHg or diastolic blood pressure >95 mmHg);

[0442] 17. History of cerebrovascular accident or myocardial infarction within 24 weeks (168 days) of treatment;

[0443] 18. History of renal failure, dialysis or renal transplantation;

[0444] 19. Known sensitivity to any compound to be administered in treatment; and / or

[0445] 20. Pregnant or breastfeeding women

[0446] Thus, the present invention includes methods of treating or preventing DR and / or DME in a subject in need thereof, wherein the subject

[0447] 1. Age ≥ 50 years old;

[0448] 2. Active subfoveal CNV;

[0449] 3. Best Corrected Visual Acuity (BCVA) Early Treatment Study for Diabetic Retinopathy

[0450] (ETDRS) letter scores ranged from approximately 78 to 24;

[0451] 4. Central retinal thickness ≥300 μm or ≥320 μm;

[0452] 5. The lesion area is about 6 or 7 mm 2 ;

[0453] 6. DRSS score better than or equal to 43, 47 or worse; and / or

[0454] 7. Suffering from type 1 or type 2 diabetes;

[0455] and / or,

[0456] 1. No signs of macular edema in the eye due to any cause other than diabetes;

[0457] 2. No IOP ≥ 25 mmHg in the eye;

[0458] 3. No history of glaucoma filtration surgery in the past, or unlikely to require filtration surgery in the eyes in the future;

[0459] 4. No signs of infectious blepharitis, keratitis, scleritis, or conjunctivitis in either eye within 4 weeks (28 days) of treatment;

[0460] 5. No intraocular inflammation and / or eye infection within 12 weeks (84 days) of treatment;

[0461] 6. No history of idiopathic or autoimmune uveitis in the eye;

[0462] 7. No vitreomacular traction or epiretinal membrane in the eye;

[0463] 8. No preretinal fibrosis involving the macula in the eye;

[0464] 9. No history of macular hole of stage 2 or above in the eye;

[0465] 10. No current iris neovascularization, vitreous hemorrhage and / or tractional retinal detachment;

[0466] 11. No history of corneal transplantation or corneal dystrophy in the eye;

[0467] 12. There are no concurrent ocular conditions that, in the opinion of the treating physician, may increase the risk to the subject beyond that anticipated from the standard IVT injection procedure, or that may otherwise interfere with the VEGF antagonist injection procedure;

[0468] 13. No other medical history, metabolic dysfunction, physical examination results, or clinical laboratory findings that reasonably suspect a disease or condition that contraindicates the use of VEGF antagonists;

[0469] 14. No previous systemic (IV) anti-VEGF administration;

[0470] 15. No uncontrolled diabetes defined by hemoglobin A1c (HbA1c) > 12%;

[0471] 16. No uncontrolled blood pressure (defined as systolic blood pressure >160 mm Hg or diastolic blood pressure >95 mm Hg);

[0472] 17. No history of cerebrovascular accident or myocardial infarction within 24 weeks (168 days) of treatment;

[0473] 18. No history of renal failure, dialysis or kidney transplantation;

[0474] 19. No known sensitivity to any compound to be administered in treatment; and / or

[0475] 20. Women who are not pregnant or breastfeeding;

[0476] The method comprises administering to an eye of the subject a single initial dose of a VEGF receptor fusion protein of about 8 mg or more (preferably aflibercept), followed by one or more second doses of the VEGF receptor fusion protein of about 8 mg or more, followed by one or more third doses of the VEGF receptor fusion protein of about 8 mg or more; wherein each second dose is administered about 2 to 4 weeks after the immediately previous dose; and wherein each third dose is administered about 12-20 weeks after the immediately previous dose.

[0477] VEGF antagonists

[0478] The present 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 a native VEGF receptor, e.g., molecules that bind to VEGF or a VEGF receptor and prevent or block the interaction between VEGF and a VEGF receptor. Specific exemplary VEGF antagonists include anti-VEGF antibodies, anti-VEGF receptor antibodies, and VEGF receptor fusion proteins. While VEGF receptor fusion proteins, such as aflibercept, are preferably used in conjunction with the methods described herein, the scope of the present invention includes methods in which any of the VEGF antagonists described herein (e.g., scFv, DARPin, anti-VEGF antibody) is used in place of such fusion proteins.

[0479] For the purposes herein, a "VEGF receptor fusion protein" refers to a molecule comprising one or more VEGF receptors or domains thereof, fused to another polypeptide that interferes with the interaction between VEGF and a native VEGF receptor, e.g., wherein two such fusion polypeptides associate 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 within the context of the present disclosure that fall within this definition include chimeric polypeptides that comprise two or more immunoglobulin (Ig)-like domains of a VEGF receptor, such as VEGFR1 (also known as Flt1) and / or VEGFR2 (also known as Flk1 or KDR), and may also contain a multimerization domain (e.g., an Fc domain).

[0480] An exemplary VEGF receptor fusion protein is the molecule designated VEGF1R2-FcΔC1(a), which is encoded by the nucleic acid sequence of SEQ ID NO: 1, or nucleotides 79 to 1374 or 79 to 1371 thereof.

[0481] VEGF1R2-FcΔC1(a) contains three components:

[0482] (1) a VEGFR1 component comprising amino acids 27 to 129 of SEQ ID NO: 2;

[0483] (2) a VEGFR2 component comprising amino acids 130 to 231 of SEQ ID NO: 2; and

[0484] (3) A multimerization component ("FcΔC1(a)") comprising amino acids 232 to 457 of SEQ ID NO:2 (the C-terminal amino acid of SEQ ID NO:2, i.e., K458, may or may not be included in VEGF receptor fusion proteins; see U.S. Patent Nos. 7,396,664 or 7,354,579, each of which is incorporated herein for all purposes). Note that amino acids 1 to 26 of SEQ ID NO:2 are a signal sequence.

[0485] If the multimerization component (MC) of the VEGF receptor fusion protein is derived from an IgG (e.g., IgG1) Fc domain, the amino acids of the MC are no less than amino acids 232 to 457 of SEQ ID NO: 2. Therefore, the IgG MC cannot be truncated to be shorter than 226 amino acids.

[0486] In one embodiment of the invention, the VEGF receptor fusion protein comprises amino acids 27-458 or 27-457 of SEQ ID NO: 2 (eg, in the form of a homodimer).

[0487]

[0488] (SEQ ID NO: 1)

[0489] MVSYWDTGVLLCALLSCLLLTGSSSG SDTGPFVEMYSEIPEIIHMTEGRELVIPCRVTS

[0490] PNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLT

[0491] HRQTNTIIDVVLSSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRD

[0492] LKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPPC

[0493] PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT

[0494] KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY

[0495] TLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK

[0496] LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0497] (SEQ ID NO: 2)

[0498] In an embodiment of the present invention, the VEGF receptor fusion protein comprises

[0499] (1) the immunoglobulin-like (Ig) domain 2 of a first VEGF receptor (e.g., VEGFR1), and

[0500] (2) Ig domain 3 of a second VEGF receptor (e.g., VEGFR2),

[0501] (3) and optionally, further comprising an Ig domain 4 of a second VEGF receptor (e.g., VEGFR2) and

[0502] (4) Multimeric components (e.g., the Fc domain of IgG, including the hinge, CH2, and CH3 domains).

[0503] For example, in an embodiment of the present invention, the VEGF receptor fusion protein has the following arrangement of domains:

[0504] [VEGFR1 Ig domain 2]-[VEGFR2 Ig domain 3]-[MC] (e.g., a homodimer thereof) or

[0505] • [VEGFR1 Ig domain 2]-[VEGFR2 Ig domain 3]-[VEGFR2 Ig domain 4]-[MC] (eg, a homodimer thereof).

[0506] Note that the present disclosure also includes within its scope high concentration formulations comprising a VEGF binding molecule or anti-VEGF antibody or antigen-binding fragment thereof or a biopolymer conjugate thereof (e.g., KSI-301) (but not a VEGF receptor fusion protein), e.g.

[0507] Bevacizumab (e.g., at a concentration of about 80-90 or 88 mg / ml),

[0508] Ranibizumab (e.g., at a concentration of about 20-40 mg / ml, such as 21-35, 21 or 35 mg / ml),

[0509] anti-VEGF aptamers, such as pegaptanib (e.g., pegaptanib sodium),

[0510] Single chain (eg V L -V H ) an anti-VEGF antibody, such as brolucizumab (e.g., at a concentration of about 200-400 or 200, 210, 400, or 420 mg / ml),

[0511] an anti-VEGF DARPin, such as Abicipar Pegol DARPin (e.g., at a concentration of about 70-140, 70, or 140 mg / ml), or

[0512] • A bispecific anti-VEGF antibody, e.g., which also binds ANG2, such as RG7716 (farximab) (e.g., at a concentration of about 100-400, 100, 105, 400, or 420 mg / ml).

[0513] To minimize repetition of the embodiments discussed herein, it is contemplated that the scope of the present invention includes embodiments in which any formulation discussed herein includes an anti-VEGF antibody or antibody fragment or other VEGF binding molecule discussed herein (e.g., substituted with an anti-VEGF DARPin) at any concentration discussed herein in place of a VEGF receptor fusion protein. For example, the present invention includes formulations containing 35 or 80 mg / ml ranibizumab, a buffer, a thermal stabilizer, a viscosity reducer, and a surfactant.

[0514] DARPins are designed ankyrin repeat proteins. DARPins typically contain three to four tightly packed repeats of approximately 33 amino acid residues, each containing a β-turn and two antiparallel α-helices. This rigid framework provides protein stability while enabling the presentation of the variable region, typically consisting of six amino acid residues per repeat, for target recognition.

[0515] An "anti-VEGF" antibody or antigen-binding fragment of an antibody is one that specifically binds to VEGF.

[0516] Exemplary VEGF receptor fusion proteins include aflibercept ( Regeneron Pharmaceuticals, Inc.) or Conbercept (sold by Chengdu Kanghong Biotechnology Co., Ltd.). See International Patent Application Publication No. WO2005 / 121176 or WO2007 / 112675. The terms "aflibercept" and "conbercept" include biosimilars thereof. A biosimilar version of a reference product (e.g., aflibercept) generally refers to a product containing the same amino acid sequence, but includes biosimilar products under the U.S. Biologics Price Competition and Innovation Act.

[0517] The present invention also includes embodiments that include administration of one or more additional therapeutic agents in addition to a VEGF antagonist, for example, administration (in one or more doses) of a second VEGF antagonist, an antibiotic, an anesthetic (e.g., a local anesthetic), a nonsteroidal anti-inflammatory drug (NSAID), a steroid (e.g., a corticosteroid, dexamethasone), triamcinolone acetonide (TA), methotrexate, rapamycin, an anti-tumor necrosis factor alpha drug (e.g., infliximab), daclizumab, and / or an inhibitor of a complement component (e.g., C3 or C5) to the eye receiving the injection.

[0518] pharmaceutical preparations

[0519] The present invention includes methods in which a VEGF antagonist administered to the eye of a subject is contained 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 "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or state government, or listed in the U.S. Pharmacopeia or other generally recognized pharmacopoeia for use in animals (more specifically, humans). The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the VEGF antagonist is administered. Numerous suitable formulations can be found in formularies known to all medicinal chemists: Remington's Pharmaceutical Sciences (15th edition, Mack Publishing Company, Easton, Pa., 1975), for example, Chapter 87 by Blaug, Seymour.

[0520] The pharmaceutical formulations used in the methods of the present invention can be "high concentration". High concentration pharmaceutical formulations of the present invention include a VEGF antagonist, such as a VEGF receptor fusion protein, at a concentration of at least 41 mg / ml, at least 80 mg / ml, at least 100 mg / ml, at least 125 mg / ml, at least 140 mg / ml, at least 150 mg / ml, at least 175 mg / ml, at least 200 mg / ml, at least 225 mg / ml, at least 250 mg / ml, or at least 275 mg / ml. "High concentration" can refer to a formulation comprising a VEGF antagonist at a concentration of 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 certain aspects, the concentration of the VEGF antagonist in the formulation is about 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 Other VEGF antagonist concentrations are contemplated herein, provided that the concentration functions according to the embodiments herein.

[0521] In an embodiment of the invention, the concentration of the pharmaceutical formulation used in the methods of the invention is one containing about 4, 6, 8, 10, 12, 14, 16, 18 or 20 mg of a VEGF receptor fusion protein (e.g., aflibercept), or any acceptable dose of such protein discussed herein in an amount of 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; 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.

[0522] The invention includes methods using any of the formulations described herein under "Exemplary Formulations" (as discussed herein), but wherein the concentration of the VEGF receptor fusion protein (e.g., aflibercept) is substituted for the concentrations described in this section ("VEGF Receptor Fusion Proteins and Other VEGF Inhibitors").

[0523] The buffer used in the pharmaceutical formulations herein that can be used in the methods of the present invention refers to a solution that resists pH changes by using an acid-base conjugate. The buffer is capable of maintaining the pH in the range of from 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 the present invention is about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7 or about 6.8. Examples of buffers included in the formulations herein include histidine-based buffers, such as histidine and histidine hydrochloride or histidine acetate. The buffer included in the formulation herein can alternatively be a phosphate-based buffer (e.g., sodium phosphate), an acetate-based buffer (e.g., sodium acetate or acetic acid) or can be a citrate-based buffer (e.g., sodium citrate or citric acid). It is also recognized that the buffer can be a mixture of the above-mentioned substances, as long as the buffer plays the role of the buffer formulation within the above-mentioned pH range. In some cases, the buffer is from about 5mM to about 25mM, or more typically, about 5mM to about 15mM. The buffer can be about 5mM, about 6mM, about 7mM, about 8mM, about 9mM, about 10mM, about 11mM, about 12mM, about 13mM, about 14mM, about 15mM, about 16mM, about 17mM, about 18mM, about 19mM, about 20mM, about 21mM, about 22mM, about 23mM, about 24mM or about 25mM.

[0524] In an embodiment of the present invention, histidine and histidine monohydrochloride are used to prepare a histidine-based buffer.

[0525] Surfactants as used herein refer to ingredients that protect higher concentrations of VEGF antagonists from various surface and interface induced stresses, such as VEGF receptor fusion proteins. Thus, surfactants can be used to limit or minimize VEGF receptor fusion protein aggregation and promote protein solubility. Suitable surfactants herein have been shown to be nonionic and may include surfactants with polyoxyethylene moieties. Such exemplary surfactants include: polysorbate 20, polysorbate 80, poloxamer 188, polyethylene glycol 3350, and mixtures thereof. The surfactant in the formulation may be about 0.02% to about 0.1% weight / volume (w / v), and more typically, about 0.02% to about 0.04% (w / v). In some cases, the surfactant is about 0.02% (w / v), about 0.03% (w / v), about 0.04% (w / v), about 0.05% (w / v), about 0.06% (w / v), about 0.07% (w / v), about 0.08% (w / v), about 0.09% (w / v), or about 0.1% (w / v).

[0526] Thermostats that can be used in the pharmaceutical formulations of the methods described herein are ingredients that provide thermal stability to the thermal denaturation of the VEGF antagonist (e.g., VEGF receptor fusion protein) and prevent the loss of potency or activity of the VEGF receptor fusion protein. Suitable thermostabilizers include sugars and can be sucrose, trehalose, sorbitol, or mannitol, or can be amino acids such as L-proline, L-arginine (e.g., L-arginine monohydrochloride), or taurine. In addition, thermostabilizers can also include substituted acrylamides or propanesulfonic acid, or can be compounds such as glycerol.

[0527] In some cases, the pharmaceutical formulations used in the methods herein include sugar and taurine, sugar and amino acid, sugar and propanesulfonic acid, sugar and taurine, glycerol and taurine, glycerol and propanesulfonic acid, amino acid and taurine, or amino acid and propanesulfonic acid. In addition, the formulations can include sugar, taurine and propanesulfonic acid, glycerol, taurine and propanesulfonic acid, and L-proline, taurine and propanesulfonic acid.

[0528] The embodiments herein may have the thermal stabilizers present alone, each independently present 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 a total combination of the foregoing concentrations. The concentration of the thermal 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).

[0529] With respect to taurine and propanesulfonic acid, in embodiments of the present invention, these thermal stabilizers may be present in the formulation in an amount of about 25 mM to about 100 mM, more typically about 50 mM to about 75 mM (compared to other thermal stabilizers).

[0530] Viscosity reducing agent is generally used for reducing or preventing protein aggregation.The viscosity reducing agent included herein includes: sodium chloride, magnesium chloride, D- or L-arginine (such as L-arginine monohydrochloride), lysine or its mixture.When being present in this article, viscosity reducing agent can be from about 10mM to about 100mM, more typically from about 30mM to about 75mM, even more typically from about 40mM to about 70mM amount exists.In some cases, viscosity reducing agent exists with the amount of about 10mM, about 15mM, about 20mM, about 25mM, about 30mM, about 35mM, about 40mM, about 45mM, about 50mM, about 55mM, about 60mM, about 65mM, about 70mM, about 75mM, about 80mM, about 85mM, about 90mM, about 95mM or about 100mM.

[0531] The pharmaceutical formulations for the methods described herein can also have a pharmaceutically acceptable viscosity for ocular administration, such as intravitreal injection. Viscosity generally refers to the measure of the resistance of a fluid to deformation by shear stress or tensile stress (usually measured by techniques known in the art, such as a viscometer or rheometer). The typical viscosity of the formulations for the methods described herein is 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. Thus, the viscosity of the formulations herein can be about 5.0 cP, about 6.0, about 7.1 cP, about 7.2 cP, about 7.3 cP, about 7.4 cP, about 7.5 cP, about 7.6 cP, about 10 cP, about 10.5 cP, about 11.0 cP, about 11.5 cP, about 12.0 cP, about 12.5 cP, about 13.0 cP, about 13.5 cP, about 14.0 cP, about 14.5 cP, or about 15.0 cP (e.g., when measured at 20°C).

[0532] Various embodiments herein do not require the inclusion of inorganic salts or other viscosity reducing agents to maintain these very useful viscosities. Typically, high concentration protein solutions require viscosity reducing agents to avoid protein aggregation and higher viscosities, which make the formulations difficult to inject intravitreally and reduce the effectiveness of VEGF receptor fusion proteins. Therefore, embodiments herein include methods using formulations that are substantially free of or do not contain added sodium chloride (NaCl), magnesium chloride (MgCl2), D- or L-arginine (e.g., L-arginine hydrochloride), lysine, or other viscosity reducing agents.

[0533] Osmolality is a key attribute of the injectable pharmaceutical formulations for the inventive method. It is hoped that the product meets physiological osmotic conditions. In addition, osmolality provides the confirmation of soluble content in the solution. In embodiments of the present invention, the osmolality of the formulation for the inventive method is less than or equal to about 506mmol / Kg or from about 250 to about 506mmol / 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 506mmol / Kg. In embodiments of the present invention, osmotic pressure is lower than about 250mmol / Kg.

[0534] Exemplary pharmaceutical formulations for use in the methods of the invention include the following:

[0535] Formulation A: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0536] Formulation B: 80 mg / ml aflibercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0537] Formulation C: 80 mg / ml aflibercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0538] Formulation D: 80 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2.

[0539] Formulation E: 80 mg / ml aflibercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0540] Formulation F: 80 mg / ml aflibercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0541] 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 a viscosity reducing agent.

[0542] Formulation H: 80 mg / ml aflibercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally specifically excluding a viscosity reducing agent.

[0543] 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 a viscosity reducing agent.

[0544] 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 specifically excluding a viscosity reducing agent.

[0545] Formulation K: 80 mg / ml aflibercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally specifically excludes viscosity reducing agents.

[0546] 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 excludes viscosity reducing agents.

[0547] Formulation M: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0548] Formulation N: 150 mg / ml aflibercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0549] Formulation O: 150 mg / ml aflibercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0550] Formulation P: 150 mg / ml aflibercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2.

[0551] Formulation Q: 150 mg / ml aflibercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0552] Formulation R: 150 mg / ml aflibercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0553] 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 a viscosity reducing agent.

[0554] Formulation T: 150 mg / ml aflibercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2 (eg, 6.2), and optionally specifically excluding a viscosity reducing agent.

[0555] 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 excludes viscosity reducing agents.

[0556] 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 a viscosity reducing agent.

[0557] Formulation W: 150 mg / ml aflibercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally specifically excluding a viscosity reducing agent.

[0558] 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 a viscosity reducing agent.

[0559] 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.

[0560] Formulation Z: 80 mg / ml Conbercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0561] Formulation AA: 80 mg / ml Conbercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0562] Formulation BB: 80 mg / ml Conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2.

[0563] Formulation CC: 80 mg / ml Conbercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0564] Formulation DD: 80 mg / ml Conbercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0565] 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 a viscosity reducing agent.

[0566] Formulation FF: 80 mg / ml Conbercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 20, with a pH of 5.8 to 6.2, and optionally specifically excluding a viscosity reducing agent.

[0567] 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 a viscosity reducing agent.

[0568] 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 excludes viscosity reducing agents.

[0569] Formulation II: 80 mg / ml Conbercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 80, pH 5.8 to 6.2, and optionally specifically excluding a viscosity reducing agent.

[0570] 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 excludes viscosity reducing agents.

[0571] Formulation KK: 150 mg / ml Conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0572] Formulation LL: 150 mg / ml Conbercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0573] Formulation MM: 150 mg / ml Conbercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0574] Formulation NN: 150 mg / ml Conbercept, 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 6.2.

[0575] Formulation OO: 150 mg / ml Conbercept, 10 mM phosphate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0576] Formulation PP: 150 mg / ml Conbercept, 10 mM citrate-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 80, and 40 mM sodium chloride, pH 5.8 to 6.2.

[0577] 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 a viscosity reducing agent.

[0578] Formulation RR: 150 mg / ml Conbercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose and 0.03% (w / v) polysorbate 20, pH 5.8 to 6.2, and optionally specifically excluding a viscosity reducing agent.

[0579] Formulation SS: 150 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 excludes viscosity reducing agents.

[0580] 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 a viscosity reducing agent.

[0581] Formulation UU: 150 mg / ml Conbercept, 10 mM phosphate-based buffer, 8% (w / v) sucrose, and 0.03% (w / v) polysorbate 80, with a pH of 5.8 to 6.2, and optionally, specifically, excluding a viscosity reducing agent.

[0582] 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 a viscosity reducing agent.

[0583] Formulation WW: 140 mg / ml VEGF receptor fusion protein (eg, aflibercept), 10 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM taurine, pH 5.8.

[0584] 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.

[0585] 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.

[0586] 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.

[0587] Formulation AAA: 140 mg / ml VEGF receptor fusion protein (eg, aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8.

[0588] Formulation BBB: 140 mg / ml VEGF receptor fusion protein (e.g., Aflibercept), 20 mM histidine-based buffer, 2.5% (w / v) sucrose, 2.0% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8.

[0589] Formulation CCC: 80, 100, 120, or 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM arginine (e.g., arginine hydrochloride), pH 5.8.

[0590] Formulation DDD: 140 mg / ml VEGF receptor fusion protein (eg, aflibercept), 10 mM histidine-based buffer, 4% (w / v) proline, 0.03% (w / v) polysorbate 20, and 50 mM PSA, pH 5.8.

[0591] Formulation EEE: 140 mg / ml VEGF receptor fusion protein (eg, 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.

[0592] Formulation FFF: 140 mg / ml VEGF receptor fusion protein (eg, aflibercept), 10 mM sodium phosphate, 5% (w / v) sucrose, and 0.03% polysorbate 20, pH 6.2.

[0593] 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

[0594] 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

[0595] 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

[0596] 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

[0597] 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

[0598] 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

[0599] 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

[0600] 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

[0601] 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)

[0602] 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)

[0603] 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)

[0604] 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)

[0605] 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)

[0606] Formulation TTT: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20 mM histidine, pH 5.8; 5% sucrose, 0.03% polysorbate 20; 100 mM L-arginine (e.g., L-arginine hydrochloride)

[0607] Formulation UUU: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0608] Formulation VVV: 30 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0609] Formulation: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0610] Formulation XXX: 60 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0611] Formulation YYY: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 10% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0612] Formulation ZZZ: 120 mg / ml VEGF receptor fusion protein (e.g., aflibercept); 20% sucrose, 10 mM phosphate, 0.03% polysorbate 20, pH 6.2

[0613] 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

[0614] 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

[0615] Formulation CCCC: 140 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 10 mM sodium phosphate, 5% sucrose, 40 mM sodium chloride, 0.03% PS20, pH 6.2

[0616] Formulation DDDD: 80 mg / ml VEGF receptor fusion protein (e.g., aflibercept), 20 mM histidine-based buffer, 5% (w / v) sucrose, 0.03% (w / v) polysorbate 20, and 50 mM L-arginine (e.g., L-arginine hydrochloride), pH 5.8.

[0617] Formulation EEEE: 120.0 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±12 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine hydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0618] Formulation FFFF: 113.3 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., 102-125 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0619] 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).

[0620] Formulation HHHH: 100.0 mg / ml VEGF receptor fusion protein (e.g., aflibercept) (e.g., ±10 mg / ml), 20 mM histidine-based buffer (e.g., ±2 mM), 5% (w / v) sucrose (e.g., ±0.5%), 0.03% (w / v) polysorbate 20 (e.g., 0.02%-0.04%), and 50 mM L-arginine (e.g., L-arginine monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0621] 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 monohydrochloride) (e.g., ±5 mM), pH 5.8 (e.g., 5.6-6.0 or 5.5-6.1).

[0622] Formulation JJJJ: 150 mg / ml aflibercept (e.g., aflibercept) (e.g., ±15 mg / ml), 10 mM sodium phosphate, 8% (w / v) sucrose (e.g., ±0.8%), 0.03% (w / v) polysorbate 20 (e.g., 0.02%-0.04%), and 50 mM L-arginine (e.g., L-arginine hydrochloride), pH 6.2 (e.g., 6.0-6.4 or 5.9-6.5).

[0623] 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).

[0624] See International Patent Application Publication No. WO2019 / 217927.

[0625] In one embodiment of the present invention, ≥8 mg of a VEGF receptor fusion protein (preferably aflibercept) is administered in an aqueous pharmaceutical formulation comprising: a VEGF receptor fusion protein comprising two polypeptides, each comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, Ig domain 3 of VEGFR2, and a multimerization component (e.g., comprising 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 sodium glutamate. In one embodiment of the present 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) an osmolarity 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 a mixture thereof. The concentration of the histidine-based buffer may be 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 species immediately after production and purification and / or less than or equal to about 6% high molecular weight species after storage at about 2°C-8°C for about 24 months.

[0626] In one embodiment of the invention, ≥8 mg of a VEGF receptor fusion protein is administered in an aqueous pharmaceutical formulation comprising: at least about 100 mg / ml of a VEGF receptor fusion protein comprising two polypeptides, each polypeptide comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, Ig domain 3 of VEGFR2, and a multimerization component (e.g., aflibercept); about 10 mM to 100 mM L-arginine; sucrose; a histidine-based buffer; and a surfactant; wherein the formulation has a pH of about 5.0 to about 6.8; 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 to 8°C for about 24 months.

[0627] In one embodiment of the present invention, the aqueous pharmaceutical formulation comprises:

[0628] ≥ about 100 mg / ml VEGF receptor fusion protein (e.g., aflibercept), histidine-based buffer, and L-arginine;

[0629] Approximately 140 mg / ml aflibercept; 20 mM histidine-based buffer; 5% sucrose; 0.03% polysorbate 20; 10 mM L-arginine; pH 5.8;

[0630] Approximately 150 ± 15 mg / ml aflibercept, 10 mM phosphate-based 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;

[0631] About 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;

[0632] 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;

[0633] About 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;

[0634] ≥ approximately 100 mg / ml aflibercept, histidine-based buffer, and L-arginine;

[0635] ≥ about 100 mg / ml aflibercept, about pH 5.8, wherein the formulation forms <3% HMW aggregates after incubation at 5°C for 2 months;

[0636] Approximately 114.3 mg / mL aflibercept; 10 mM-50 mM histidine-based buffer, sugar, nonionic surfactant, L-arginine, pH 5.8;

[0637] or

[0638] 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.

[0639] In one embodiment of the present invention, ≥8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation when administered, the aqueous pharmaceutical formulation comprising

[0640] 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);

[0641] a heat stabilizer which is a sugar, an amino acid, sucrose, mannitol, sorbitol, trehalose, L-proline, glycine, glycerol, taurine, or propanesulfonic acid (e.g., about 2% (w / v) to about 10% (w / v), e.g., 5% (w / v));

[0642] a buffer which is a histidine-based buffer, a phosphate-based buffer, an acetate-based buffer (e.g., at a concentration of about 5-25 mM, such as 10 mM or 20 mM); or a citrate-based buffer;

[0643] a nonionic surfactant, such as polyoxyethylene-based polysorbate 20, polysorbate 80, poloxamer 188, or polyethylene glycol 3350 (e.g., at a concentration of about 0.02% to about 0.1% (w / v), e.g., 0.03% (w / v)); and

[0644] a viscosity reducing agent, which is NaCl, MgCl2, D-arginine, L-arginine or L-lysine (e.g., at a concentration of about 10-100 mM, such as 50 mM),

[0645] wherein the pH of the formulation is from about 5.0 to about 6.8 (eg, 5.0-6.0 or 5.8).

[0646] In one embodiment of the invention, the concentration of aflibercept in the aqueous pharmaceutical formulation is about 100 mg / ml; 101 mg / ml; 102 mg / ml; 103 mg / ml; 104 mg / ml; 105 mg / ml; 106 mg / ml; 107 mg / ml; 108 mg / ml; 109 mg / ml; 110 mg / ml; 111 mg / ml; 112 mg / ml; 113 mg / ml; 113.3 mg / ml; 114 mg / ml; 114.1 mg / ml; 114.2 mg / ml; 114.3 mg / ml; 114.4 mg / ml; 114.5 mg / ml; 114.6 mg / ml; 114.7 mg / ml; 114.8mg / 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 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. ;

[0647] 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 at a pH of about 5.0 to about 6.8; wherein 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 2-8°C for about 24 months.

[0648] 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; and the pH is about 5.0 to about 6.8.

[0649] Treatment and administration

[0650] The present invention provides methods for treating angiogenic eye diseases (e.g., DR and / or DME) by 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, e.g., a VEGF receptor fusion protein such as 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 angiogenic eye diseases such as diabetic macular edema (DME) and / or diabetic retinopathy (DR) by sequentially administering one or more (e.g., 3 or 4 or 5) doses of about 8 mg or more of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) about every 2-4 weeks or 3-5 weeks, e.g., monthly (or about every 28 days, 28±5 days or about every 4 weeks), followed by one or more doses of about 8 mg or more of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) every 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. A dosing regimen comprising a 12-week third dosing interval may be referred to herein as a 12-week dosing regimen or 8q12 or HDq12; a dosing regimen comprising a 16-week third dosing interval may be referred to herein as a 16-week dosing regimen or 8q16 or HDq16; and a dosing regimen comprising a 20-week third dosing interval may be referred to herein as a 20-week dosing regimen or 8q20 or HDq20.

[0651] Additionally, the present invention includes methods for treating angiogenic eye diseases (e.g., DR and / or DME) by administering ≥8 mg of a VEGF receptor fusion protein, preferably aflibercept, one or more times every 4 weeks, 8 weeks, 12-20 weeks, 12-16 weeks, 12 weeks, or 16 weeks; and the first 3, 4, or 5 doses are administered every 4 weeks, followed by dosing approximately every 8 weeks.

[0652] In one embodiment of the invention, after the 8 mg monthly loading dose, the subject begins receiving a maintenance dose of ≥ 8 mg every 12 weeks, 16 weeks, or 20 weeks without any intermediate doses. The subject enters the maintenance dose phase quickly / immediately after the loading dose phase. In one embodiment of the invention, the subject continues to receive ≥ 8 mg for 12 weeks, 16 weeks, or 20 weeks without any intermediate doses.

[0653] In one embodiment of the invention, the subject has not received a dosing regimen modification (DRM) or discontinued treatment for at least 1, 2, 3, 4, or 5 years.

[0654] The present invention also provides a method for improving vision in a subject with type 1 or type 2 diabetes (e.g., a subject with diabetic macular edema or diabetic retinopathy) by sequentially administering one or more (e.g., 3 or 4 or 5) doses about monthly (or about every 28 days, 28±5 days, or about every 4 weeks), followed by one or more doses every 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.

[0655] The terms "initial dose," "secondary dose," and "tertiary dose" refer to the temporal order in which a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) is administered. Thus, the "initial dose" is the dose administered at the beginning of a treatment regimen (also referred to as the "baseline amount"); the "secondary dose" is the dose administered after the initial dose; and the "tertiary dose" is the dose administered after the second dose. The initial, second, and third doses can all contain the same amount of VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), but typically differ from one another in the frequency of administration. However, in certain 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 one another during the course of treatment (e.g., adjusted upward or downward, as appropriate).

[0656] Therefore, the dosage regimen of the present invention can be expressed as follows:

[0657] A method for treating an angiogenic eye disease (e.g., DME or DR) in a subject in need thereof, the method comprising administering (e.g., intravitreally administering) to a subject in need thereof

[0658] about ≥ 8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, such as about 50 μl; 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 74 μl; 75 μl or 100 μl) of a single initial dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0659] One or more (e.g., 2 or 3 or 4, preferably 2) second doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0660] one or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept);

[0661] wherein each second dose is administered 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose; and

[0662] wherein each third dose is administered 12 or 16 or 20 weeks (preferably about 12-16, 12 or 16 or 20 weeks) after the immediately preceding dose.

[0663] The present invention includes such methods wherein, in addition to any scheduled initial, second, and / or third dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), one or more additional, non-scheduled doses are administered to a subject. Such doses are typically administered at the discretion of the treating physician based on the particular needs of the subject. The present invention also provides methods for treating angiogenic eye diseases (e.g., DR or DME) by administering to a subject in need thereof about ≥8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, such as about 50 μl; 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 74 μl; 75 μl; 76 μl; 77 μl; 78 μl; 79 μl; 80 μl; 81 μl; 82 μl; 83 μl; 84 μl; 85 μl; 86 μl; 87 μl; 88 μl; 89 μl; 90 μl; 91 μl; 92 μl; 93 μl; 94 μl; 95 μl; 96 μl; 97 μl; 98 μl; 99 μl; 100 μl or less, or 100 μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept).

[0664] An on-demand (PRN) treatment regimen requires that physician visits be maintained at fixed intervals (e.g., every 2, 3, 4, 8, 12, 16, or 20 weeks), with the decision to administer an injection of a VEGF receptor fusion protein based on the anatomic findings at each respective visit. An upper-limited PRN dosing regimen is a PRN in which the subject must be treated at a certain minimum frequency, e.g., at least once every 2, 3, or 4 months.

[0665] The treat and extend (T&E) regimen requires adjusting the intervals between doctor 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 without any leakage), the next interval or intervals can be extended; if there is fluid accumulation, the next interval will be shortened. An injection of the VEGF receptor fusion protein will be given at each visit after the T&E; the current clinical status only affects the duration of the next injection interval.

[0666] The present invention includes embodiments in which at any point during the HDq12-20, HDq12, HDq16, or HDq20 treatment regimen, a patient can be switched to a PRN, capped PRN, or T&E regimen. PRN, capped PRN, and / or T&E can continue indefinitely or can be discontinued at any point and then restarted at any stage of the HDq12-20, HDq12, HDq16, or HDq20 regimen. Any HDq12-20, HDq12, HDq16, or HDq20 regimen can be preceded or followed by a period of PRN, capped PRN, and / or T&E.

[0667] The invention encompasses methods comprising administering a desired dose of a HDq12 or HDq16 regimen, wherein each third dose is administered 12 or 16 weeks after the immediately 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), for example, until signs of disease activity or visual impairment worsen or recur, and then continuing dosing during the last third interval used or the second to last third interval used.

[0668] The invention encompasses methods comprising administering a regimen of HDq12-20 or HDq12 or HDq16 or HDq20 at the desired doses, wherein the treatment interval between any two third doses is reduced (e.g., from 20 weeks to 19, 18, 17 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, as a result of improvement in signs of disease activity or visual impairment (e.g., BCVA stabilizes or improves and / or CRT stabilizes or decreases), and thus, optionally, the dosage interval can be extended, for example, back to a greater interval length.

[0669] For example, in one embodiment of the invention, the dosage interval, for example during a 12-week or 16-week dosing period, can be extended, for example by 4-week intervals as appropriate (e.g., from 12 weeks to 16 weeks or 16 weeks to 20 weeks), for example if:

[0670] For example, BCVA loss of < 5 letters from week 12; and / or

[0671] CRT <300 μm on SD-OCT (or <320 μm as determined by Spectralis SD-OCT).

[0672] In one embodiment of the invention, the subject receives an initial, second interval, followed by a third interval of 12 or 16 weeks, and then after about 1 year, the third interval is extended to about 20 weeks.

[0673] In one embodiment of the invention, the method of treating angiogenic eye diseases such as DR or DME as described herein comprises the step of evaluating BCVA and / or CRT, and if one or both criteria are met, extending the interval as discussed.

[0674] For example, in one embodiment of the invention, the dosage interval, e.g., during a dosing period of 12-20 weeks or 12 weeks or 16 weeks or 20 weeks, can be shortened (e.g., from 12 weeks or 16 weeks or 20 weeks to 8 weeks; from 16 weeks or 20 weeks to 12 weeks or from 20 weeks to 16 weeks), e.g., if:

[0675] For example, a BCVA (ETDRS or Snellen equivalent) loss of greater than 5 or 10 letters (e.g., relative to the BCVA observed at about 12 or 16 weeks after initiation of treatment), e.g., due to persistent or worsening DR or DME; and / or

[0676] • An increase in CRT greater than 25 or 50 microns is observed (e.g., relative to the CRT observed approximately 12 weeks after initiation of treatment).

[0677] In one embodiment of the invention, if a subject receiving the HDq12 regimen meets the criteria for a reduced dosing interval, e.g., at week 16 or 20, the dosing interval is reduced to 8 weeks. In one embodiment of the invention, if a subject receiving the HDq16 regimen meets the criteria for a reduced dosing interval, e.g., at week 16 or 20, the dosing interval is reduced to 8 weeks; and if a subject receiving the HDq16 regimen meets the criteria for a reduced dosing interval, e.g., at week 24, the dosing interval is reduced to 12 weeks. In one embodiment of the invention, the interval is not reduced to any interval shorter than 8 weeks. In one embodiment of the invention, the method of treating angiogenic eye diseases such as DR or DME as described herein includes the step of evaluating BCVA and / or CRT, and if one or both criteria are met, the interval is shortened as discussed.

[0678] See also Figure 3 or Figure 4 .

[0679] The present invention also provides a method for treating angiogenic eye diseases (preferably DME or DR) by administering:

[0680] about ≥8 mg dose (e.g., 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; 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), approximately once every 4, 5, 6, 7, 8, 12, 16 or 20 weeks;

[0681] or

[0682] A single initial dose (e.g., 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; or 100 μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0683] One or more (e.g., 2 or 3 or 4 (preferably 2 or 4)) second doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by

[0684] one or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept);

[0685] wherein each second dose is administered 2 to 4 weeks (preferably 4 weeks) after the immediately preceding dose; and wherein each third dose is administered about 4, 5, 6, 7 or 8 weeks (e.g., 8 weeks) after the immediately preceding dose;

[0686] or

[0687] about ≥ 8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, such as about 50 μl; 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl; 72 μl; 73 μl; 74 μl; 75 μl ; 76 μl; 77 μl; 78 μl; 79 μl; 80 μl; 81 μl; 82 μl; 83 μl; 84 μl; 85 μl; 86 μl; 87 μl; 88 μl; 89 μl; 90 μl; 91 μl; 92 μl; 93 μl; 94 μl; 95 μl; 96 μl; 97 μl; 98 μl; 99 μl; or 100 μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) approximately every 4 weeks (q4w);

[0688] or

[0689] less than about 8 or 9 doses (e.g., about 5 doses or 6 doses) of about ≥ 8 mg (e.g., about 100 μl or less, about 75 μl or less, or about 70 μl or less, e.g., about 50 μl; 51 μl; 52 μl; 53 μl; 54 μl; 55 μl; 56 μl; 57 μl; 58 μl; 59 μl; 60 μl; 61 μl; 62 μl; 63 μl; 64 μl; 65 μl; 66 μl; 67 μl; 68 μl; 69 μl; 70 μl; 71 μl) over the course of about 48 weeks or 100 μl) of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept).

[0690] Dosing every "month" or "one month" refers to dosing after about 28 days, about 4 weeks, or about 28 ± 5 days and can cover up to 5 weeks ± 5 days. Dosing every "4 weeks" or "4 weeks" refers to dosing after about 28 days (± 5 days), about one month, or about 28 days (± 5 days), and can cover up to every 5 weeks (± 5 days).

[0691] Dosing every "2-4 weeks" or "2-4 weeks" refers to dosing after about 2 weeks (± 5 days), 3 weeks (± 5 days), or 4 weeks (± 5 days). Dosing every "8 weeks" or "8 weeks" refers to dosing after about 2 months (± 5 days) or about 56 (± 5 days).

[0692] Dosing every "12 weeks" or after "12 weeks" refers to dosing after about 3 months, about 84 days (± 5 days), about 90 days (± 5 days), or about 84 (± 5 days). Dosing every "16 weeks" or after "16 weeks" refers to dosing after about 4 months or about 112 days (± 5 days).

[0693] Administration every "12-20 weeks" or administration after "12-20 weeks" refers to administration after 12, 13, 14, 15, 16, 17, 18, 19 or 20 weeks (±5 days), preferably after about 12-16 weeks (±5 days), about 12 weeks (±5 days), about 16 weeks (±5 days) or about 20 weeks (±5 days).

[0694] Administration every "12-20 weeks" refers to administration after about 12, 13, 14, 15, 16, 17, 18, 19 or 20 weeks (±5 days), preferably after about 12-16 weeks (±5 days), about 12 weeks (±5 days), about 16 weeks (±5 days) or about 20 weeks (±5 days).

[0695] A dose of ≥8 mg encompasses doses of about 8 mg or doses above 8 mg, for example about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg.

[0696] In embodiments of the invention, any dosing frequency specified herein can 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.

[0697] "Sequential administration" means that each dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) is administered to the subject's eye at a different time point, for example, on a different day separated by a predetermined time interval (e.g., hours, days, weeks, or months). The present invention includes methods comprising sequentially administering to the subject's eye a single initial dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by one or more second doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept), followed by one or more third doses of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept).

[0698] An effective or therapeutically effective dose of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) for treating or preventing an angiogenic eye disease refers to an amount of a VEGF antagonist (e.g., a VEGF receptor fusion protein such as aflibercept) sufficient to alleviate one or more signs and / or symptoms of the disease or condition in the subject being treated, whether by inducing 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 about ≥8 mg three times per month, followed by once every 12-20 weeks. In one embodiment of the invention, reduction in signs and / or symptoms is, for example, achieving an increase in BCVA of ≥5, 10, or 15 letters (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 a BCVA of ≥69 letters; achieving the absence of fluid in the center of the fovea; a reduction in central retinal thickness (CRT) of about 150 microns or more (e.g., less than 300 microns in nAMD subjects / patients; and / or a reduction of at least about 200 microns in DR or RVO patients / subjects) or achieving a normal CRT (e.g., about 300 microns or less); and / or achieving no leakage on fluorescein angiography.

[0699] Baseline values ​​refer to values ​​before the start of treatment (before drug administration).

[0700] "Angiogenic eye disease" refers to any eye disease caused by or associated with blood vessel growth or proliferation or vascular leakage. Non-limiting examples of angiogenic eye diseases treatable or preventable using the methods of the present invention include:

[0701] Age-related macular degeneration (neovascular AMD),

[0702] Macular edema (ME),

[0703] Macular edema after retinal vein occlusion (ME-RVO),

[0704] Retinal vein occlusion (RVO),

[0705] Central retinal vein occlusion (CRVO),

[0706] Branch retinal vein occlusion (BRVO),

[0707] Diabetic macular edema (DME),

[0708] Choroidal neovascularization (CNV),

[0709] Iris neovascularization,

[0710] Neovascular glaucoma,

[0711] Fibrosis after glaucoma surgery,

[0712] Proliferative vitreoretinopathy (PVR),

[0713] Optic disc neovascularization,

[0714] Corneal neovascularization,

[0715] · Retinal neovascularization,

[0716] Vitreous neovascularization,

[0717] Pannus,

[0718] Pterygium,

[0719] Vascular retinopathy,

[0720] Diabetic retinopathy (DR) (e.g., non-proliferative diabetic retinopathy (e.g., characterized by a Diabetic Retinopathy Severity Scale (DRSS) level of about 47 or 53) or proliferative diabetic retinopathy; e.g., in a subject who does not have DME), and

[0721] Diabetic retinopathy in subjects with diabetic macular edema (DME).

[0722] The present invention provides methods for treating angiogenic eye diseases (e.g., DR and / or DME) in a subject in need thereof by 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, e.g., a VEGF receptor fusion protein such as aflibercept) (e.g., about every 2-4 weeks or 3-5 weeks, preferably every 4 weeks; preferably three initial loading doses), followed by additional doses every 12-20 weeks, preferably 12-16 weeks, 12 weeks, 16 weeks, or 20 weeks, wherein the subject achieves and / or maintains, e.g., at week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 after initiation of treatment:

[0723] Improvement in the Diabetic Retinopathy Severity Scale (DRSS), e.g., at least grade 2 or 3;

[0724] Improvement in best-corrected visual acuity;

[0725] Dry retina

[0726] Improvement in best-corrected visual acuity;

[0727] BCVA improved by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 letters or ≥5, ≥10, or ≥15 letters;

[0728] A BCVA of at least 69 letters;

[0729] A decrease in central retinal thickness (CRT), for example, by about 100, 125, 150, 175, or 200 microns;

[0730] Absence of vascular leakage as measured by fluorescein angiography (FA);

[0731] Improvement in the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) total score compared to pre-treatment baseline;

[0732] The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea and in central subregions;

[0733] Maintaining a fluid-free retina (total fluid, IRF, and / or SRF in the foveal center and central subregion);

[0734] No macular edema;

[0735] Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT); and / or

[0736] Do not leave the HDq12 or HDq16 treatment regimen once started.

[0737] In one embodiment of the invention, a subject receiving HDq12 or HDq16 or HDq20 treatment as described herein for angiogenic eye diseases (e.g., DR and / or DME) achieves one or more of the following:

[0738] not received a dose regimen modification, e.g., where the dose interval (e.g., the interval of the third dose) was decreased once initiated from a HDq12-20 or HDq12 or HDq16 or HDq20 treatment regimen, e.g., by at least 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks;

[0739] receiving 100% of all scheduled doses, for example, at least 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96 weeks;

[0740] Non-inferiority of BVCA compared with intravitreal aflibercept 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, followed by 2 mg approximately every 8 weeks or every 2 months;

[0741] an increase in BCVA (based on ETDRS letter score) of about 7, 8, or 9 letters by Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96, for example, where baseline BCVA is about 61, 62, or 63;

[0742] BCVA improvement 4 weeks after treatment initiation of approximately 4 or 5 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 4 or 5 letters (ETDRS or Snellen equivalent) with HDq16;

[0743] BCVA improvement 8 weeks after treatment initiation of approximately 6 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0744] BCVA improvement 12 weeks after treatment initiation of approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0745] BCVA improvement 16 weeks after treatment initiation was approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq12, or 7 letters (ETDRS or Snellen equivalent) with HDq16;

[0746] Improvement in BCVA 20 weeks after treatment initiation of approximately 6 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0747] Improvement in BCVA 24 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with HDq16;

[0748] Improvement in BCVA 28 weeks after treatment initiation of approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0749] Improvement in BCVA 32 weeks after treatment initiation was approximately 7 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0750] Improvement in BCVA 36 weeks after treatment initiation of 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16

[0751] Improvement in BCVA 40 weeks after treatment initiation was approximately 8 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16;

[0752] Improvement in BCVA 44 weeks after treatment initiation was approximately 8 letters (ETDRS or Snellen equivalent) with HDq12, or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16;

[0753] An improvement in BCVA 48 weeks after the initiation of treatment of about 8 or 9 letters (ETDRS or Snellen equivalents) on HDq12 or about 7 or 8 letters (ETDRS or Snellen equivalents) on HDq16, for example, where baseline BCVA is about 61, 62, or 63 letters (ETDRS or Snellen equivalents);

[0754] An improvement in BCVA 60 weeks after the initiation of treatment of about 8 or 9 letters (ETDRS or Snellen equivalents) on HDq12 or about 7 or 8 letters (ETDRS or Snellen equivalents) on HDq16, for example, where baseline BCVA is about 61, 62, or 63 letters (ETDRS or Snellen equivalents);

[0755] An improvement in BCVA of about 8 or 9 letters, or up to 40 letters (ETDRS or Snellen equivalents) from Week 48 to Week 60 on the HDq12 regimen, e.g., when baseline BCVA is about 63 or 64; or an improvement in BCVA of about 7 or 8 letters, or up to 40 letters (ETDRS or Snellen equivalents) from Week 48 to Week 60 on the HDq16 regimen, e.g., when baseline BCVA is about 61 or 62;

[0756] Improvement in BCVA at about Week 8 after the initiation of treatment that is maintained thereafter (e.g., within about ±1 or ±2 ETDRS letters or Snellen equivalents) during the treatment regimen, e.g., to at least Week 48;

[0757] Improvement in best-corrected visual acuity (based on ETDRS letter score) (e.g., from the start of treatment to weeks 12, 24, 36, 48, 60, 72, 84, 90, or 96);

[0758] Improvement in best-corrected visual acuity (BVCA) from the start of treatment to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, or Week 48;

[0759] An increase in BCVA (e.g., from the start of treatment to Weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48) of ≥4 letters, ≥5 letters, ≥6 letters, ≥7 letters, ≥8 letters, ≥9 letters, or ≥10 letters, as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or Snellen equivalent;

[0760] a change in BCVA score (based on ETDRS letter score) of about 7, 8, or 9 from the start of treatment between Week 36 and Week 48, for example, wherein the BCVA at any point between Week 36 and Week 48 is about 60 or 70;

[0761] a change in BCVA score (based on ETDRS letter scores) of up to 38 letters from the start of treatment between Week 36 and Week 48, when using the HDq12 or HDq16 regimen, e.g., where BCVA at baseline is between about 27 and 79;

[0762] a change in BCVA score (based on ETDRS letter score) from the start of treatment of about 7, 8, or 9 between Week 48 and Week 60; for example, wherein the BCVA at any point between Week 48 and Week 60 is about 69, 70, 71, 72, or 73;

[0763] When the baseline BCVA is approximately 73 ETDRS letters or less on the HDq12 regimen, for example, an improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0764] When the baseline BCVA is about >73 ETDRS letters on the HDq12 regimen, for example, an improvement in BCVA of about 5 or 6 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0765] When the baseline BCVA is approximately 73 ETDRS letters or less on the HDq16 regimen, for example, an improvement in BCVA of approximately 8 or 9 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0766] When the baseline BCVA is approximately >73 ETDRS letters on the HDq16 regimen, for example, an improvement in BCVA of approximately 4 or 5 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0767] When taking the HDq12 regimen, when the baseline CRT is < about 400 microns, for example, an improvement in BCVA of about 7 or 8 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0768] When using the HDq12 regimen, when baseline CRT is ≥400 microns, for example, BCVA improvement of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after treatment initiation;

[0769] When taking HDq16, when the baseline CRT is < about 400 microns, for example, an improvement in BCVA of about 5 or 6 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0770] When taking HDq16, when the baseline CRT is ≥ about 400 microns, for example, an improvement in BCVA of about 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment;

[0771] No loss of 5, 10, or 15 letters (based on the ETDRS letter score) by Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96;

[0772] An improvement of 5, 10, or 15 letters (based on the ETDRS letter score) by week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96;

[0773] Have not lost 5, 10, 15, or 69 letters or more of BCVA (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 after the start of treatment);

[0774] BCVA score (based on ETDRS letter score) of approximately 69, 70, 71, 72, or 73 between weeks 48 and 60;

[0775] BCVA (based on ETDRS letter scores) of at least about 69 letters, for example, by week 48 or week 60;

[0776] BCVA 4 weeks after the start of treatment is approximately 68 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0777] BCVA 8 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0778] BCVA 12 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0779] BCVA 16 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0780] BCVA 20 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0781] BCVA 24 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0782] BCVA 28 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0783] BCVA 32 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0784] BCVA 36 weeks after the start of treatment of approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0785] BCVA 40 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0786] BCVA 44 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0787] BCVA 48 weeks after the start of treatment is approximately 73 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen;

[0788] BCVA of about 71, 72, 73, or 74 letters (ETDRS or Snellen equivalents) from Week 36 to Week 48 on the HDq12 regimen, e.g., when baseline BCVA is about 57, 58, 59, 60, 61, 62, 63, or 64; or BCVA of about 69, 70, 71, 72, or 73 (ETDRS or Snellen equivalents) from Week 36 to Week 48 on the HDq16 regimen, e.g., when baseline BCVA is about 55, 56, 57, 58, 59, 60, 61, or 62;

[0789] BCVA of about 69 or 70 or up to 94 (ETDRS or Snellen equivalents) from Week 48 to Week 60 on the HDq12 regimen, e.g., when baseline BCVA is about 63 or 64; or BCVA of about 72 or 73 or up to 89 (ETDRS or Snellen equivalents) from Week 48 to Week 60 on the HDq16 regimen, e.g., when baseline BCVA is about 61 or 62;

[0790] Improvement in BCVA of ≥5, ≥10, or ≥15 letters (based on the ETDRS letter score) (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96);

[0791] Diabetic Retinopathy Severity Scale (DRSS) improvement of ≥ 2 grades (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 72, 84, 90, or 96);

[0792] Diabetic Retinopathy Severity Scale (DRSS) improvement of ≥ 3 grades (e.g., from the start of treatment to Weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 72, 84, 90, or 96);

[0793] The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) in the center or central subregion of the fovea (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96) (e.g., as measured by optical coherence tomography (OCT));

[0794] No vascular leakage in the retina as measured by fluorescein angiography (FA) (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);

[0795] Maintenance of retinal fluid-free (total fluid, IRF, and / or SRF in the foveal center and central subregion) (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96);

[0796] Total fluorescein leakage area within the ETDRS grid at week 48 or week 60 (mm 2 ) is reduced by approximately 12, 12.6, 13, 13.6, 13.9 or 14 mm 2 or more, or up to about 57 or 68mm 2 (e.g., measured by fluorescein angiography);

[0797] When using the HDq12 regimen, at week 48, the total area of ​​fluorescein leakage within the ETDRS grid (mm 2 ) is reduced by approximately 13, 13.3, 13.9 or 14 mm 2 or more (e.g., up to about 52 mm 2 )(e.g., measured by fluorescein angiography);

[0798] When using the HDq16 regimen, at week 48, the total area of ​​fluorescein leakage within the ETDRS grid (mm 2 ) is reduced by approximately 7, 7.7, 8, 9, 9.4 or 10 mm 2 or more (e.g., up to about 55 mm 2 )(e.g., measured by fluorescein angiography);

[0799] Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT) (e.g., from the start of treatment to weeks 12, 24, 36, 48, 60, 72, 84, 90, or 96);

[0800] The retina has no fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea (e.g., from the start of treatment to weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48);

[0801] retinal dryness (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96);

[0802] Absence of central foveal fluid (e.g., from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, or 96) (e.g., as measured by optical coherence tomography (OCT));

[0803] Change in central retinal thickness four weeks after the start of treatment was approximately -118 or -118.3 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -124, -125, -124.9, or -125.5 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0804] The change in central retinal thickness at 8 weeks after the start of treatment was approximately -137 or -137.4 microns (±17, 18, or 19 microns) with HDq12; or approximately -139, -140, -139.6, or -140.3 microns (±17, 18, or 19 microns) with HDq16;

[0805] Change in central retinal thickness 12 weeks after the start of treatment was approximately -150 or -150.1 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -152, -153, -152.7, or -153.4 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0806] The change in central retinal thickness 16 weeks after the start of treatment was approximately -139 or -139.4 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -145, -146, -145.5, or -146.4 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0807] Change in central retinal thickness 20 weeks after the start of treatment was approximately -117 or -117.1 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -112, -113, -112.5, or -113.3 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0808] Change in central retinal thickness 24 weeks after initiation of treatment was approximately -158 or -158.1 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -103, -104, -103.8, or -104.3 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0809] Change in central retinal thickness 28 weeks after the start of treatment was approximately -146, -147, or -146.7 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -162 or -162.3 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0810] The change in central retinal thickness at 32 weeks after the start of treatment was approximately -132 microns (±17, 18, or 19 microns) with HDq12; or approximately -145, -146, or -145.8 microns (±17, 18, or 19 microns) with HDq16;

[0811] Change in central retinal thickness 36 weeks after initiation of treatment was approximately -168 or -168.1 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -124, -125, -124.7, or -125.2 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0812] The change in central retinal thickness 40 weeks after the start of treatment was approximately -163 microns (±17, 18, or 19 microns) with HDq12; or approximately -122, -123, -122.5, or -123.1 microns (±17, 18, or 19 microns) with HDq16;

[0813] Change in central retinal thickness 44 weeks after initiation of treatment was approximately -147, -148, or -147.4 microns (±17, 18, or 19 microns) on the HDq12 regimen; or approximately -164, -164.1, or -164.3 microns (±17, 18, or 19 microns) on the HDq16 regimen;

[0814] the change in central retinal thickness 48 weeks after the initiation of treatment is about -171 or -172 or -171.7, -172, -173, -174, -175, -176 or -176.77 microns (± 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 microns) on the HDq12 regimen; or about -148 or -149 or -148.3 or -149.4 microns (± 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 microns) on the HDq16 regimen, for example, where the baseline CRT is about 449, 450, 455 or 460 microns;

[0815] The change in central retinal thickness at 60 weeks after the initiation of treatment was approximately -181, -182, -181.95, -176, -176.24, or -177 (±6, 10, 17, 18, or 19 microns) on the HDq12 regimen (e.g., where the baseline CRT was approximately 460 microns); or approximately -166, -166.26, -167, or -167.18 microns (±8, 9, 10, 17, 18, or 19 microns) on the HDq16 regimen (e.g., where the baseline CRT was approximately 457 microns);

[0816] The change in central retinal thickness between the start of treatment (week 0) and week 4 on the HDq12 regimen was approximately -118, -119, or -118.3 microns;

[0817] A change in central retinal thickness of approximately -19, -20, or -19.1 microns between weeks 4 and 8 on the HDq12 regimen;

[0818] A change in central retinal thickness of approximately -12, -13, or -12.7 microns between weeks 8 and 12 on the HDq12 regimen;

[0819] A change in central retinal thickness of approximately -40 or -41 microns between weeks 20 and 24 on the HDq12 regimen;

[0820] A change in central retinal thickness of approximately -36, -37, or -36.1 microns between weeks 32 and 36 on the HDq12 regimen;

[0821] A change in central retinal thickness of approximately -24, -25, or -24.3 microns between weeks 44 and 48 on the HDq12 regimen;

[0822] A change in central retinal thickness of approximately -124, -125, or -124.9 microns between the start of treatment (week 0) and week 4 on the HDq16 regimen;

[0823] A change in central retinal thickness of approximately -14, -15, or -14.7 microns between weeks 4 and 8 on the HDq16 regimen;

[0824] A change in central retinal thickness of approximately -13, -14, or -13.1 microns between weeks 8 and 12 on the HDq16 regimen;

[0825] A change in central retinal thickness of approximately -58, -59, or -58.5 microns between weeks 24 and 28 on the HDq16 regimen;

[0826] A change in central retinal thickness of approximately -41, -42, or -41.6 microns between weeks 40 and 44 on the HDq16 regimen;

[0827] a reduction in CRT at week 4, 5, 6, 7, or 8 after initiation of treatment that is maintained thereafter (e.g., within about ±17, ±18, or ±19 microns) during the treatment regimen, e.g., through at least week 48;

[0828] a decrease in central retinal thickness (CRT) of approximately 100, 125, 150, 175, or 200 microns (e.g., from the start of treatment to weeks 12, 24, 36, 48, 60, 72, 84, 90, or 96);

[0829] a decrease in CRT by about 148-182 microns (e.g., 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183 microns) (e.g., as measured by optical coherence tomography (OCT)) by week 48 or week 60, for example, where the baseline CRT is about 449, 450, 455, or 460 microns;

[0830] a decrease in central retinal thickness (CRT), e.g., by at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 microns (e.g., from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48);

[0831] ocular safety (e.g., intraocular pressure) and non-ocular safety (e.g., hypertensive events or APTC events) or mortality in subjects with angiogenic eye diseases such as DR or DME was similar to that of aflibercept given intravitreally at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections, followed by 2 mg intravitreally approximately every 8 weeks or every 2 months;

[0832] the free aflibercept concentration in plasma is about 0.149 (±0.249) mg / l at about 0.1667 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0833] the free aflibercept concentration in plasma is about 0.205 (±0.250) mg / l at about 0.3333 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0834] The free aflibercept concentration in plasma is about 0.266 (±0.211) mg / l about 1 day after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0835] The free aflibercept concentration in plasma is about 0.218 (±0.145) mg / l about 2 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0836] The free aflibercept concentration in plasma is about 0.140 (±0.0741) mg / l about 4 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0837] the free aflibercept concentration in plasma is about 0.0767 (±0.0436) mg / l at about 7 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0838] the free aflibercept concentration in plasma is about 0.0309 (±0.0241) mg / l at about 14 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0839] the free aflibercept concentration in plasma is about 0.0171 (±0.0171) mg / l at about 21 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0840] The free aflibercept concentration in plasma is about 0.00730 (±0.0113) mg / l at about 28 days after the first dose; for example, wherein the free aflibercept concentration in plasma is undetectable at baseline, for example, wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0841] an adjusted bound aflibercept concentration in plasma of about 0.00698 (±0.0276) mg / l at about 0.1667 days after the first dose; for example, wherein at baseline, there was an adjusted bound aflibercept concentration of about 0.00583 mg / l (±0.0280), and wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0842] an adjusted bound aflibercept concentration in plasma of about 0.00731 (±0.0279) mg / l at about 0.3333 days after the first dose; for example, wherein at baseline, there was an adjusted bound aflibercept concentration of about 0.00583 mg / l (±0.0280), and wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0843] an adjusted bound aflibercept concentration in plasma of about 0.0678 (±0.0486) mg / l at about 1 day after the first dose; for example, wherein at baseline, there was an adjusted bound aflibercept concentration of about 0.00583 mg / l (±0.0280), and wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0844] An adjusted bound aflibercept concentration in plasma of about 0.138 (±0.0618) mg / l about 2 days after the first dose; for example, wherein at baseline, there was an adjusted bound aflibercept concentration of about 0.00583 mg / l (±0.0280), and wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0845] an adjusted bound aflibercept concentration in plasma of about 0.259 (±0.126) mg / l at about 4 days after the first dose; for example, wherein at baseline, there was an adjusted bound aflibercept concentration of about 0.00583 mg / l (±0.0280), and wherein the subject has not received intravitreal VEGF inhibitor (e.g., aflibercept) treatment for at least 12 weeks;

[0846] an adjusted bound aflibercept concentration in plasma of about 0.346 (±0.151) mg / l at about 7 days after the...

Claims

1. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 is 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 ● After the intravitreal injection of aflibercept, the time to reach the lower limit of quantification (LLOQ) of free aflibercept in the subject's plasma 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 1-2, 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 1-3, 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 5-6, 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 5-7, 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 5-8, 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 10-11, 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 10-12, 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 10-13, 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 1-14, wherein <4 mg is about 2 mg or 2-4 mg.

16. The method of any one of claims 1-15, wherein the subject suffers from diabetic retinopathy and / or diabetic macular edema.

17. The method of any one of claims 1-16, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising a histidine-based buffer.

18. The method of any one of claims 1-17, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising arginine.

19. The method of any one of claims 1-18, 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 1-19, 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 1-20, wherein the >8 mg aflibercept is in an aqueous pharmaceutical formulation comprising sucrose.

22. The method of any one of claims 1-21, 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 1-22, 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 - Treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, - Improve the best corrected visual acuity in subjects in need with DR and / or DME; or - Promote retinal dryness in subjects in need with DR and / or DME; 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 1-24, for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to the eye of the subject A single initial dose of about 8 mg or more of 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 1-24, for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, 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, 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 1-24, for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to the eye of the subject A single initial dose of about 8 mg or more of 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 1-24, for treating or preventing diabetic retinopathy (DR) and / or diabetic macular edema (DME) in a subject in need thereof, comprising administering to the eye of the subject A single initial dose of about 8 mg or more of a VEGF receptor fusion protein, 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.

29. A method for treating or preventing diabetic macular edema 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 8-16 weeks (2-4 months + / - 7 days).

30. A method for treating or preventing diabetic retinopathy (DR) 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 8-16 weeks (2-4 months + / - 7 days).

31. A method for treating or preventing diabetic macular edema 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).

32. A method for treating or preventing diabetic retinopathy (DR) 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).

33. A method for treating or preventing diabetic macular edema 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).

34. A method for treating or preventing diabetic retinopathy (DR) 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).

35. A method for treating or preventing diabetic macular edema 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 (2-4 months + / - 7 days).

36. A method for treating or preventing diabetic retinopathy (DR) 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 (2-4 months + / - 7 days).

37. A method for treating or preventing diabetic macular edema (DME) or diabetic retinopathy (DR) in a subject in need thereof with a dose of 8 mg of aflibercept (equivalent to 70 microliters of injection), the method comprising administering to the subject 1 injection per month (every 4 weeks) for 3 consecutive doses of the 8 mg aflibercept; and then administering injections of the 8 mg aflibercept at intervals that can be extended to every 16 weeks based on the physician's judgment of visual and / or anatomical results.

38. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof: (1) wherein the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, and then the method comprises administering to the subject the initial dose of 8 mg of VEGF receptor fusion protein after 1 month, and administering the first second dose of 8 mg of VEGF receptor fusion protein 1 month thereafter; and administering the second second dose of 8 mg of VEGF receptor fusion protein 1 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) wherein 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) wherein the subject has received an initial dose of 2 mg of VEGF receptor fusion protein, and the method 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) wherein 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) wherein 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) wherein 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 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) wherein 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 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; (8) wherein 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, 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; (9) wherein 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; and 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) wherein 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, 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; 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) wherein 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, 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 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; (12) wherein 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) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject the initial 8 mg dose 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; and 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) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject 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; 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) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein one month later; 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) wherein 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 third 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; (17) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months later and 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 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; (18) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein two months later and a second 8 mg second dose of VEGF receptor fusion protein one month later; 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; (19) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later; thereafter, the method then comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein two months later and thereafter, 12 weeks, 16 weeks or 20 weeks later, administering a maintenance dose of 8 mg VEGF receptor fusion protein once or more for 12 weeks or 16 weeks once a week according to the HDq12, HDq16 or HDq20 dosing regimen; (20) wherein 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 received a third second dose of 2 mg of VEGF receptor fusion protein one month later and received a fourth second dose of 2 mg of VEGF receptor fusion protein one month later; Thereafter, the method then comprises administering to the subject a first 8 mg maintenance dose of the VEGF receptor fusion protein after 2 months, and administering all other 8 mg maintenance doses of the VEGF receptor fusion protein every 12 weeks, 16 weeks, or 20 weeks according to the HDq12, HDq16, or HDq20 dosing regimen; (21) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and received one or more 2 mg maintenance doses every 8 weeks thereafter; then the method comprises administering to the subject the initial 8 mg dose of VEGF receptor fusion protein two months after the last VEGF receptor fusion protein maintenance dose 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; (22) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter; then the method comprises administering to the subject a first 8 mg second dose of VEGF receptor fusion protein two months after the last VEGF receptor fusion protein maintenance dose and 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; (23) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a second 8 mg second dose of VEGF receptor fusion protein two months after the last maintenance dose of VEGF receptor fusion protein and thereafter administering a maintenance dose of 8 mg VEGF receptor fusion protein once or more times for 12 weeks, 16 weeks or 20 weeks thereafter according to the HDq12, HDq16 or HDq20 dosing regimen; or (24) wherein 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 received a second 2 mg second dose of VEGF receptor fusion protein one month later and received a third 2 mg second dose of VEGF receptor fusion protein one month later and received a fourth 2 mg second dose of VEGF receptor fusion protein one month later and has received one or more 2 mg maintenance doses every 8 weeks thereafter, then the method comprises administering to the subject a first 8 mg maintenance dose of VEGF receptor fusion protein two months after the last VEGF receptor fusion protein maintenance dose, 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.

39. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, said subject having received a dosing regimen for treating or preventing said condition, wherein: (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; or (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 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; or (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 1 month later and received a second 8 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 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 schedule; or (d) wherein 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 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 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.

40. A method for treating or preventing an angiogenic eye disease in a subject in need thereof, said subject having received a dosing regimen for treating or preventing said condition, said dosing regimen requiring: 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.

41. The method of any one of claims 1-40, wherein the subject has received a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema: A single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by 4 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.

42. The method of any one of claims 1-41 for treating or preventing diabetic retinopathy or diabetic macular edema 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 wherein each third dose is administered about 12 weeks or 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.

43. The method of claim 42, wherein during the treatment period, the subject exhibits (a) BCVA loss < 5 letters; and / or (b) CRT < 300 or 320 μm.

44. The method of any one of claims 42-43, 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.

45. The method of any one of claims 1-45, for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 wherein each third dose is administered about 12 weeks, 16 weeks, or 20 weeks after the immediately preceding dose; The method further comprises shortening the third dose interval 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.

46. ​​The method of claim 45, wherein during the treatment period, the subject exhibits (a) BCVA loss >10 letters from baseline; and / or (b) CRT increase relative to baseline >50 μm.

47. The method of any one of claims 45-46, 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.

48. The method of any one of claims 1-47, 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 reduced from 12 or 16 weeks to 8 weeks; or if the interval is 24 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 reduced from 16 weeks to 12 weeks.

49. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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; wherein after the 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.

50. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema 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 two 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 the 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 said VEGF receptor fusion protein at said reduced or extended dose intervals, wherein Criteria for extending the interval include:

1. BCVA loss < 5 letters; and / or 2. CRT < 300 or 320 microns; And, wherein the criteria for reducing the interval include:

1. BCVA loss > 10 letters; 2. Persistent or worsening DME; and / or 3. CRT increases > 50 microns.

51. The method of claim 50, wherein the criteria for extending the interval include:

1. BCVA loss < 5 letters from week 12; as well as 2. CRT measured by SD-OCT < 300 or 320 μm; and / or wherein the criteria for reducing said interval include both of the following:

1. BCVA loss >10 letters combined with persistent or worsening DME since week 12; and 2. CRT increase > 50 microns from week 12 onwards.

52. The method of any one of claims 49-51, wherein if the criteria are met, the interval is extended to 12 weeks, 16 weeks or 20 weeks.

53. A method for treating or preventing diabetic retinopathy and / or diabetic macular edema in a subject in need thereof, the subject having 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 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 Each third dose is administered approximately 12-20 weeks after the immediately preceding dose.

54. A method for treating or preventing an angiogenic eye disease in a subject in need thereof, the method comprising administering to an 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.

55. The method of claim 54, wherein the angiogenic eye disease is diabetic retinopathy and / or diabetic macular edema.

56. The method of any one of claims 54-55, wherein one or more second doses is 2, 3 or 4 second doses and / or about 2-4 weeks is about 4 weeks.

57. The method of any one of claims 1-56, 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 administering a VEGF receptor fusion protein to said eye.

58. The method of claim 57, comprising the steps of: 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 are found in the subject, the subject is excluded from the administration.

59. The method of any one of claims 1-58, further comprising monitoring the subject for conjunctival hemorrhage, cataracts, vitreous detachment, vitreous floaters, corneal epithelial defects, and / or increased intraocular pressure during the treatment or prevention period.

60. The method of any one of claims 1-59, comprising providing, prior to each administration, 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 tip and a bevel; ·One No. 30 x 1 / 2-inch injection needle; and A 1-mL Luer-lock syringe with a graduation marking 70 μL capacity; packaged together; then (1) visually inspecting the aqueous formulation in the vial and, if particulates, turbidity, or discoloration are visible, using another vial of the aqueous formulation containing the VEGF receptor fusion protein; (2) removing 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 the 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 until the plunger tip is aligned with the 70 μl mark on the syringe.

61. The method of any one of claims 1-60, wherein 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.

62. The method of any one of claims 1-61, wherein the subject has been receiving a dosing regimen for treating or preventing diabetic retinopathy and / or diabetic macular edema that requires: A single initial dose of about 2 mg of a VEGF receptor fusion protein, followed by four 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.

63. The method of any one of claims 1 to 62, 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 is 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 the one or more second doses is 2 second doses, and the VEGF receptor fusion protein is aflibercept.

64. The method of any one of claims 1-63, wherein the VEGF receptor fusion protein comprises amino acids 27-457 of the amino acid sequence shown in SEQ ID NO:

2.

65. The method of any one of claims 1-64, wherein the VEGF receptor fusion protein is selected from aflibercept and conbercept.

66. The method of any one of claims 1-65, wherein the VEGF receptor fusion protein: (i) comprises 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) comprises two polypeptides comprising immunoglobulin-like (Ig) domain 2 of VEGFR1, Ig domain 3 of VEGFR2, Ig ​​domain 4 of VEGFR2, and a multimerization component; or (iv) comprising two VEGFR1R2-FcΔC1(a) polypeptides, wherein the two VEGFR1R2-FcΔC1(a) polypeptides are encoded by the nucleic acid sequence of SEQ ID NO:

1.

67. The method of claim 66, wherein the VEGF receptor fusion protein comprises two polypeptides comprising the immunoglobulin-like (Ig) domain 2 of VEGFR1, the Ig domain 3 of VEGFR2, and a multimerization component.

68. The method of any one of claims 1-67, wherein the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation selected from the group consisting of A-KKKK.

69. The method of any one of claims 1-68, wherein the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 114.3 mg / ml VEGF receptor fusion protein.

70. The method of any one of claims 1-69, comprising administering the VEGF receptor fusion protein to both eyes of the subject.

71. The method of any one of claims 1-70, wherein the VEGF receptor fusion protein is administered from a pre-filled syringe.

72. The method of claim 71, wherein the prefilled syringe is glass or plastic, and / or sterile.

73. The method of any one of claims 1-72, wherein the VEGF receptor fusion protein is injected intravitreally using a 30 gauge x 1 / 2 inch sterile injection needle.

74. The method of any one of claims 1-73, wherein the subject has previously received one or more doses of 2 mg VEGF receptor fusion protein.

75. The method of any one of claims 1-74, wherein one or more additional doses of the VEGF receptor fusion protein are administered.

76. The method of any one of claims 1-74, wherein 2 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising 40 mg / ml of VEGF receptor fusion protein.

77. The method of claim 76, wherein 2 mg of the 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.

78. The method of any one of claims 1-77, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising a sugar or a polyol.

79. The method of any one of claims 1-78, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising sucrose.

80. The method of any one of claims 1-79, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation at a pH of about 5.

8.

81. The method of any one of claims 1-80, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 103-126 mg / ml of the VEGF receptor fusion protein, a histidine-based buffer, and arginine.

82. The method of any one of claims 1-81, wherein 8 mg of the VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising about 114.3 mg / ml of the VEGF receptor fusion protein, a histidine-based buffer, and arginine.

83. The method of any one of claims 1-82, 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.

84. The method of any one of claims 1-83, 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.

85. The method of claims 1-84, wherein 8 mg of VEGF receptor fusion protein is in an aqueous pharmaceutical formulation comprising >100 mg / ml VEGF receptor fusion protein, histidine-based buffer, and L-arginine; 140 mg / ml Aflibercept; 20 mM histidine-based buffer; 5% sucrose; 0.03% polysorbate 20; 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.

86. The method of any one of claims 1-85, wherein the 8 mg of VEGF receptor fusion protein is dissolved 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, 69μl, 70μl, 71μl, 72μl, 73μl, 74μl, 75μl, 76μl, 77μl, 78μl, 79μl, 80μl, 81μl, 82μl, 83μl, 84μl, 85μl l, administered in volumes of 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.

87. The method of claim 86, wherein the VEGF receptor fusion protein is administered in a volume of about 70 ± 4 or 5 microliters.

88. The method of any one of claims 1-87, comprising administering the VEGF receptor fusion protein to both eyes of the subject.

89. The method of any one of claims 1-88, wherein the subject achieves and / or maintains one or more of: Improvement in the Diabetic Retinopathy Severity Scale (DRSS); Improvement in best corrected visual acuity; Dry retina ● Improvement in best corrected visual acuity; ●BCVA of at least 69 letters; ●No fluid in the center of the fovea; ●Decrease in central retinal thickness (CRT); ● Absence of vascular leakage as measured by fluorescein angiography (FA); ●National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score improved compared to pre-treatment baseline; The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea and in central subregions; ●Maintaining the retina free of fluid (total fluid, IRF, and / or SRF in the foveal center and central subregions); ●No macular edema; ● Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT); and / or ● Not coming off of the HDq12 or HDq16 treatment regimen once initiated.

90. The method of any one of claims 1-89, wherein the subject achieves and / or maintains one or more of the following: Non-inferiority of BVCA compared with intravitreal aflibercept at 2 mg approximately every 4 weeks for the first 3, 4, or 5 injections followed by 2 mg approximately every 8 weeks or every 2 months; An increase in BCVA (based on ETDRS letter scores) of approximately 7, 8, or 9 letters from the start of treatment to Week 60, where baseline BCVA is approximately 61, 62, or 63; BCVA (based on ETDRS letter scores) of at least about 69 letters from the start of treatment to Week 48 or Week 60; No loss of 5, 10, 15, or 69 letters or more of BCVA after 12, 24, 36, 48, 60, 72, 84, 90, or 96 weeks from the start of treatment; Improvement in BCVA (based on ETDRS letter score) from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Improvement in BVCA from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; BCVA score (based on ETDRS letter score) of approximately 69, 70, 71, 72, or 73 between weeks 48 and 60; A change in BCVA score (based on ETDRS letter score) from the start of treatment of about 7, 8, or 9 between Week 36 and Week 48, with a BCVA of about 60 or 70 at any point between Week 36 and Week 48; A change in BCVA score (based on ETDRS letter score) from the start of treatment of about 7, 8, or 9 between Week 48 and Week 60, with a BCVA of about 69, 70, 71, 72, or 73 at any point between Week 48 and Week 60; An increase in BCVA as measured by the Early Treatment Diabetic Retinopathy Study (ETDRS) eye chart or Snellen equivalent of ≥4 letters, ≥5 letters, ≥6 letters, ≥7 letters, ≥8 letters, >9 letters, or >10 letters from the start of treatment to Week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; No loss of 5, 10, or 15 letters (based on the ETDRS letter score) from the start of treatment to week 48 or week 60; An improvement of 5, 10, or 15 letters (based on ETDRS letter score) from the start of treatment to week 48 or week 60; BCVA improvement 4 weeks after treatment initiation of approximately 4 or 5 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 4 or 5 letters (ETDRS or Snellen equivalent) on HDq16; BCVA improvement 8 weeks after treatment initiation of approximately 6 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) on HDq16; BCVA improvement 12 weeks after treatment initiation of approximately 6 or 7 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16; BCVA improvement 16 weeks after treatment initiation of approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq12 or 7 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 20 weeks after initiation of treatment of approximately 6 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 6 letters (ETDRS or Snellen equivalent) on HDq16; Improvement in BCVA 24 weeks after initiation of treatment of approximately 7 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 5 or 6 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 28 weeks after initiation of treatment of approximately 7 or 8 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) on HDq16; Improvement in BCVA 32 weeks after initiation of treatment of approximately 7 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 36 weeks after initiation of treatment of 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 40 weeks after initiation of treatment of approximately 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 6 or 7 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 44 weeks after initiation of treatment of approximately 8 letters (ETDRS or Snellen equivalent) with HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) with HDq16; Improvement in BCVA 48 weeks after initiation of treatment of approximately 8 or 9 letters (ETDRS or Snellen equivalent) on HDq12 or approximately 7 or 8 letters (ETDRS or Snellen equivalent) on HDq16; Improvement in BCVA at approximately week 8 after the start of treatment and thereafter maintained to at least week 48 during the treatment regimen; Improvement in BCVA score (based on ETDRS letter score) of up to 38 letters from the start of treatment between Week 36 and Week 48 when using the HDq12 or HDq16 regimen; BCVA 4 weeks after the start of treatment is approximately 68 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 66 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 8 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 12 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 16 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 20 weeks after the start of treatment is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 24 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or 67 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 28 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 32 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA at 36 weeks after the start of treatment is approximately 71 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or approximately 68 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 40 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or 69 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 44 weeks after the start of treatment is approximately 72 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA 48 weeks after the start of treatment is approximately 73 letters (ETDRS or Snellen equivalent) on the HDq12 regimen; or BCVA is approximately 70 letters (ETDRS or Snellen equivalent) on the HDq16 regimen; BCVA between Week 36 and Week 48 of about 71, 72, 73, or 74 (ETDRS or Snellen equivalents) on the HDq12 regimen; or BCVA between Week 36 and Week 48 of about 69, 70, 71, 72, or 73 (ETDRS or Snellen equivalents) on the HDq16 regimen; When taking HDq12, when the baseline BCVA is approximately <73 ETDRS letters, the BCVA improvement at week 48 after the start of treatment is approximately 9 or 10 letters (ETDRS or Snellen equivalent); When taking HDq12, when the baseline BCVA is approximately >73 ETDRS letters, the BCVA improvement at week 48 after the start of treatment is approximately 5 or 6 letters (ETDRS or Snellen equivalent); When taking HDq16, when the baseline BCVA is approximately <73 ETDRS letters, the BCVA improvement at week 48 after the start of treatment is approximately 8 or 9 letters (ETDRS or Snellen equivalent); When taking HDq16, when the baseline BCVA is approximately >73 ETDRS letters, the BCVA improvement at week 48 after the start of treatment is approximately 4 or 5 letters (ETDRS or Snellen equivalent); An improvement in BCVA of about 7 or 8 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is about < about 400 microns on the HDq12 regimen; An improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is approximately >400 microns on HDq12; An improvement in BCVA of about 5 or 6 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is about < about 400 microns on the HDq16 regimen; An improvement in BCVA of approximately 9 or 10 letters (ETDRS or Snellen equivalent) at week 48 after the start of treatment when the baseline CRT is > approximately 400 microns on the HDq16 regimen; BCVA (based on ETDRS letter score) increased by >5, >10, or ≥15 letters from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Improvement of ≥2 or >3 grades on the Diabetic Retinopathy Severity Scale (DRSS) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Improvement of ≥ 2 grades on the Diabetic Retinopathy Severity Scale (DRSS) from the start of treatment to Week 4, Week 8, Week 12, Week 16, Week 20, Week 24, Week 28, Week 32, Week 36, Week 40, Week 44, or Week 48; The retina was free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea and in the central subregions as measured by optical coherence tomography (OCT) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; No vascular leakage as measured by fluorescein angiography (FA) from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Maintenance of retinal fluid-freeness (total fluid, IRF, and / or SRF in the foveal center and central subregion) from the start of treatment to Week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Total area of ​​fluorescein leakage within the ETDRS grid (mm) measured by fluorescein angiography at week 48 or 60 2 ) is reduced by about 12, 13 or 14 mm 2 or more; Absence of retinal fluid based on spectral domain optical coherence tomography (SD-OCT) from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; The retina is free of fluid (total fluid, intraretinal fluid [IRF], and / or subretinal fluid [SRF]) at the center of the fovea from the start of treatment to Weeks 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; Retinal dryness from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; Absence of central foveal fluid as measured by optical coherence tomography (OCT) from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; The change in central retinal thickness 4 weeks after the start of treatment was about -118 or -118.3 microns on the HDq12 regimen; or about -124 or -125 or -124.9 or -125.5 microns on the HDq16 regimen; The change in central retinal thickness at 8 weeks after the start of treatment was about -137 or -137.4 microns on the HDq12 regimen; or about -139 or -140 or -139.6 or -140.3 microns on the HDq16 regimen; The change in central retinal thickness 12 weeks after the start of treatment was about -150 or -150.1 microns on the HDq12 regimen; or about -152 or -153 or -152.7 or -153.4 microns on the HDq16 regimen; The change in central retinal thickness 16 weeks after the start of treatment was about -139 or -139.4 microns on the HDq12 regimen; or about -145 or -146 or -145.5 or -146.4 microns on the HDq16 regimen; The change in central retinal thickness 20 weeks after the start of treatment was about -117 or -117.1 microns on the HDq12 regimen; or about -112 or -113 or -112.5 or -113.3 microns on the HDq16 regimen; The change in central retinal thickness 24 weeks after the start of treatment was about -158 or -158.1 microns on the HDq12 regimen; or about -103 or -104 or -103.8 or -104.3 microns on the HDq16 regimen; The change in central retinal thickness 28 weeks after the start of treatment was about -146 or -147 or -146.7 microns on the HDq12 regimen; or about -162 or -162.3 microns on the HDq16 regimen; The change in central retinal thickness at 32 weeks after the start of treatment was about -132 microns on the HDq12 regimen; or about -145, -146, or -145.8 microns on the HDq16 regimen; The change in central retinal thickness at 36 weeks after the start of treatment was about -168 or -168.1 microns on the HDq12 regimen; or about -124 or -125 or -124.7 or -125.2 microns on the HDq16 regimen; The change in central retinal thickness 40 weeks after the start of treatment was about -163 microns on the HDq12 regimen; or about -122, -123, -122.5, or -123.1 microns on the HDq16 regimen; The change in central retinal thickness at 44 weeks after the start of treatment was about -147, -148, or -147.4 microns on the HDq12 regimen; or about -164, -164.1, or -164.3 microns on the HDq16 regimen; The change in central retinal thickness at 48 weeks after the start of treatment was about -171 or -172 or -171.7 microns on the HDq12 regimen; or about -148 or -149 or -148.3 or -149.4 microns on the HDq16 regimen; The change in central retinal thickness at 60 weeks after the start of treatment was approximately -181.95 or -176.24 microns on the HDq12 regimen; or approximately -166.26 or -167.18 microns on the HDq16 regimen; The change in central retinal thickness between the start of treatment (week 0) and week 4 on the HDq12 regimen was approximately -118 or -119 or -118.3 microns; A change in central retinal thickness of approximately -19, -20, or -19.1 microns between weeks 4 and 8 on the HDq12 regimen; A change in central retinal thickness of approximately -12, -13, or -12.7 microns between weeks 8 and 12 on the HDq12 regimen; The change in central retinal thickness was approximately -40 or -41 microns between weeks 20 and 24 on the HDq12 regimen; A change in central retinal thickness of approximately -36, -37, or -36.1 microns between Week 32 and Week 36 on the HDq12 regimen; A change in central retinal thickness of approximately -24, -25, or -24.3 microns between Week 44 and Week 48 on the HDq12 regimen; A change in central retinal thickness of -4, -5, or -4.5 microns between weeks 48 and 60 on the HDq12 regimen; - Change in central retinal thickness of approximately -124, -125, or -124.9 microns between the start of treatment (week 0) and week 4 on the HDq16 regimen; A change in central retinal thickness of approximately -14, -15, or -14.7 microns between weeks 4 and 8 on the HDq16 regimen; A change in central retinal thickness of approximately -13, -14, or -13.1 microns between weeks 8 and 12 on the HDq16 regimen; A change in central retinal thickness of approximately -58, -59, or -58.5 microns between weeks 24 and 28 on the HDq16 regimen; A change in central retinal thickness of approximately -41, -42, or -41.6 microns between weeks 40 and 44 on the HDq16 regimen; A decrease in central retinal thickness at Week 4, 5, 6, 7, or 8 after the start of treatment that is thereafter maintained within approximately ±17, ±18, or ±19 microns during the treatment regimen and through at least Week 48 after the start of treatment; A decrease in central retinal thickness of approximately 100, 125, 150, 175, or 200 microns from the start of treatment to week 12, 24, 36, 48, 60, 72, 84, 90, or 96; a reduction in central retinal thickness of about 148-182 microns as measured by optical coherence tomography (OCT) at about week 48 or 60 after the start of treatment, wherein the baseline CRT is about 449, 450, 455, or 460 microns; a decrease in central retinal thickness (CRT) of at least about 100, 125, 130, 135, 140, 145, 149, 150, 155, 160, 165, 170, 171, 172, 173, 174, or 175 microns from the start of treatment to week 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48; about 0.149 (± 0.249) mg / l free aflibercept in plasma at about 0.1667 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 0.205 (± 0.250) mg / l free aflibercept in plasma at about 0.3333 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.266 (± 0.211) mg / l free aflibercept in plasma at about 1 day after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.218 (± 0.145) mg / l free aflibercept in plasma at about 2 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.140 (±0.0741) mg / l free aflibercept in plasma at about 4 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.0767 (± 0.0436) mg / l free aflibercept in plasma at about 7 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; - free aflibercept in plasma was about 0.0309 (± 0.0241) mg / l at about 14 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; about 0.0171 (±0.0171) mg / l free aflibercept in plasma at about 21 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; about 0.00730 (±0.0113) mg / l free aflibercept in plasma at about 28 days after the first dose; wherein free aflibercept was undetectable in plasma at baseline, and wherein the subject has not received intravitreal aflibercept treatment for at least 12 weeks; At approximately 0.1667 days after the first dose, the adjusted bound aflibercept in plasma was approximately 0.00698 (±0.0276) mg / l; wherein at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - adjusted bound aflibercept in plasma was about 0.00731 (± 0.0279) mg / l at about 0.3333 days after the first dose; wherein at baseline, there was about 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.0678 (±0.0486) mg / l adjusted bound aflibercept in plasma at approximately 1 day after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (±0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.138 (± 0.0618) mg / l adjusted bound aflibercept in plasma at approximately 2 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.259 (± 0.126) mg / l adjusted bound aflibercept in plasma at approximately 4 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.346 (± 0.151) mg / l adjusted bound aflibercept in plasma at approximately 7 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.374 (± 0.110) mg / l adjusted bound aflibercept in plasma at approximately 14 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.343 (± 0.128) mg / l adjusted bound aflibercept in plasma at approximately 21 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; - approximately 0.269 (± 0.149) mg / l adjusted bound aflibercept in plasma at approximately 28 days after the first dose; wherein at baseline, there was approximately 0.00583 mg / l (± 0.0280) adjusted bound aflibercept, wherein the subject had not received intravitreal aflibercept treatment for at least 12 weeks; The maximum level of free aflibercept in plasma was reached approximately 0.965 days after the first dose; The free aflibercept in the plasma reached a maximum level of approximately 0.310 mg / l (± 0.263); The free aflibercept in the plasma is about 0 to about 1.08 mg / L; The maximum free aflibercept in the plasma (mg / l) per dose (mg) of aflibercept was approximately 0.388 (±0.0328) mg / l / mg; The adjusted combined aflibercept levels in plasma reached their maximum level approximately 14 days after the first dose; The adjusted combined aflibercept reached a maximum level of approximately 0.387 mg / l (± 0.135) in plasma; The adjusted bound aflibercept in the plasma is about 0.137 to about 0.774 mg / L; The adjusted maximum amount of bound aflibercept in the plasma (mg / l) per dose (mg) of aflibercept was approximately 0.483 (±0.0168) mg / l / mg; No anti-drug antibodies to aflibercept after 48 or 60 weeks of treatment; Improvement in National Eye Institute Visual Function Questionnaire (NEI-VFQ) total score from pre-treatment baseline; and / or No macular edema.

91. The method of claims 89-90, wherein the dry retina lacks intraretinal fluid and / or subretinal fluid.

92. The method of any one of claims 89-91, 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.

93. The method of any one of claims 89-92, wherein 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, or 60 weeks from the start of treatment is about 48 weeks or 60 weeks from the start of treatment.

94. The method of any one of claims 1 to 93, wherein ● administering 1 initial dose, 2 secondary doses, and 3 tertiary doses of >8 mg VEGF receptor fusion protein to the subject during the first year; ● wherein said subject is administered 1 initial dose, 2 secondary doses, and 2 tertiary doses of said >8 mg VEGF receptor fusion protein during the first year; or - wherein said subject is administered 1 initial dose, 2 secondary doses, and 3 tertiary doses of said >8 mg VEGF receptor fusion protein in the first year, followed by 2-4 tertiary doses in the second year.

95. The method of any one of claims 1-94, wherein the dosage interval is adjusted (increased / maintained / decreased) based on visual and / or anatomical results.

96. The method of any one of claims 1-95, further comprising one or more cycles of as needed (PRN), capped PRN, or treat and extend (T&E) dosing.

97. The method of any one of claims 1-96, wherein the VEGF receptor fusion protein is aflibercept.

98. 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 of VEGF receptor fusion protein, or wherein the container comprises >114.3 mg / mL of VEGF receptor fusion protein, wherein the instructions include instructions for administering aflibercept to patients with DME and / or DR, The instructions include instructions for starting treatment with 8 mg of aflibercept with a monthly injection (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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