Office Action 1: Non-Final Rejection (2024-12-16)
Rejection types: Double Patenting

CLAIMS:
What is Claimed is:
1\. A formulation comprising: (a) 40 to 200 mg/ml of aflibercept produced in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to said cell culture media, wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof; (b) one or more pharmaceutically acceptable excipients selected from the group of buffering agents, preservatives, detergents and combinations thereof; and (c) said formulation having a pH between 5.0 and 8.0.
2\. The formulation of claim 1, wherein said oxidized amino acid residue is histidine.
3\. The formulation of claim 1, wherein said oxidized amino acid residue is tryptophan.
4\. The formulation of claim 2, wherein said one or more excipients includes 5-40 mM of a buffer.
5\. The formulation of claim 3, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer.
6\. The formulation of claim 2, wherein the pH is between 5.5 and 7.0.
7\. The formulation of claim 3, wherein the pH is between 5.5 and 7.0.
8\. The formulation of claim 2, further comprising 0.01% to 5.0% of an organic co-solvent.
9\. The formulation of claim 3, further comprising 0.01% to 5.0% of an organic co-solvent.
10\. The formulation of claim 2, further comprising 30 mM to 150 mM of a tonicity agent.
11\. The formulation of claim 3, further comprising 30 mM to 150 mM of a tonicity agent.
12\. The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise less than 1.0% of the total amount of aflibercept.
13\. The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise between 0.05% and 1.0% of the total amount of aflibercept.
14\. The formulation of claim 3, wherein said oxidized tryptophan variants of aflibercept comprise less than 1.0% of the total amount of aflibercept.
15\. The formulation of claim 3, wherein said oxidized tryptophan variants of aflibercept comprise between 0.2% and 1.0% of the total amount of aflibercept.
16\. The formulation of claim 1, wherein said formulation is stored under an inert gas.
17\. The formulation of claim 1, wherein said formulation is stored in a vial.
18\. The formulation of claim 1, wherein said formulation is stored in a syringe.
19\. The formulation of claim 18, wherein said formulation in said syringe is an autoinjector syringe.
20\. A method of administering a pharmaceutically effective amount of aflibercept by intravitreal injection wherein said aflibercept is made in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to said cell culture media, and wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof and one or more pharmaceutically acceptable excipients.
21\. The method of claim 20, wherein the pharmaceutically effective amount of aflibercept is 2 mg.
22\. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection once every 4 weeks.
23\. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection once every 4 weeks for the first 3 months, followed by 2 mg via intravitreal injection once every 8 weeks.
24\. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection every 4 weeks for the first 5 injections followed by 2 mg via intravitreal injection once every 8 weeks.
25\. The method of claim 20, further comprising the step of administering an anesthesia, an antiseptic and/or an antibiotic.
26\. A method of treating Neovascular (Wet) Age-Related Macular Degeneration (AMD), Macular Edema Following Retinal Vein Occlusion (RVO), Diabetic Macular Edema (DME), or Diabetic Retinopathy with aflibercept wherein said aflibercept is made in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to said cell culture media, and wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof.
27\. The method claim 26, further comprising a reduction in mean retinal thickness.
28\. The method claim 26, further comprising an increase in visual acuity.
29\. The method of claim 26, further comprising a ≥2 step improvement on an Early Treatment of Diabetic Retinopathy Scale—Diabetic Retinopathy Severity Scale (ETDRS-DRSS) from baseline.
30\. The method claim 26, further comprising a stabilization or elimination of one or more symptoms or indicia of Neovascular (Wet) Age-Related Macular Degeneration (AMD), Macular Edema Following Retinal Vein Occlusion (RVO), Diabetic Macular Edema (DME), or Diabetic Retinopathy by any clinically measurable degree.

OFFICE ACTION:
DETAILED ACTION
Notice of Pre-AIA  or AIA  Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Application
The entry into the TrackOne program as of 19 November 2024 is acknowledged.
The amendment of 16 October 2024 is entered.
Claims 1-30 are pending and are being examined on the merits.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA  as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
1. Claims 1-15, 18-28, and 30 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 13, 17, 20-25, and 28 of U.S. Patent No. 11,958,894 B2. Although the claims at issue are not identical, they are not patentably distinct from each other because the ‘894 claims overlapping compositions.
The ‘894 patent claims a prefilled syringe containing aflibercept produced in CDM, wherein the aflibercept includes oxidized variants having oxidized His or Trp residues (see e.g. claim 13). The ‘894 patent further claims inclusion of excipients, a pH of 5.5-7.5 and 40-250 mg/ml of aflibercept (see e.g. claims 17 and 22). This overlaps with the formulation as instantly claimed.
With respect to claims 2 and 3, as discussed above the ‘894 patent claims both His and Trp oxidation events.
With respect to claims 4 and 5, the ‘894 patent claims 5-40 mM of buffer (see e.g. claim 23).	With respect to claims 6 and 7, as set forth above a pH of 5.5-7.0 is claimed by ‘894.
With respect to claims 8 and 9, the ‘894 patent claims organic co-solvents at 0.01-5% (see e.g. claim 24).
With respect to claims 10 and 11, the ‘894 patent claims 30-150 mM of a tonicity agent (see e.g. claim 25).
With respect to claims 12-15, the ‘894 patent claims oxidized variants at less than 1% of the total amount of aflibercept (see e.g. claims 20 and 21).
With respect to claim 18, as discussed above ‘894 claims a syringe.
With respect to claim 19, the ‘894 patent claims an autoinjector (see e.g. claim 28).
With respect to claim 20, the ‘894 patent does not claim a method of administration. However, the Federal Circuit has found that lack of an explicit claim to a method of use does not preclude a double patenting rejection if an identical use to an identical composition is disclosed in the reference patent:
A claim to a method of using a composition is not patentably distinct from an earlier claim to the identical composition in a patent disclosing the identical use (Sun Pharmaceutical. Industries, Ltd.  v. Eli Lilly & Co., 611 F.3d 1381 (Fed. Cir. 2010)).
The ‘894 discloses administration by intravital injection (see e.g. Col. 5 lines 36-40).
With respect to claims 21-24, the ‘894 application discloses administration methods that utilizes various amounts down to 2 mg as well as different injection times (see e.g. Col.97 to Col.98).
With respect to claim 25, the ‘894 application discloses administration with an anesthesia, an antibiotic, or antiseptic (see e.g. Col.97).
With respect to claim 26, again ‘894 discloses an overlapping method (see e.g. Col.10).
With respect to claims 27, 28, and 30, the ‘894 application discloses reduction in mean retinal thickness and improvement in visual acuity, as well as stabilization or elimination of symptoms or indicia (see e.g. Col.97).
2. Claims 1-30 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-30 of U.S. Patent No. 11,542,317 B1. Although the claims at issue are not identical, they are not patentably distinct from each other because the ‘317 patent claims an overlapping formulation.
The ‘317 patent claims a formulation comprising 40-200 mg/ml aflibercept produced in CDM, wherein the aflibercept contains oxidized amino acids selected from Trp, His, or combinations thereof, an excipient is present, and the formulation has a pH of 5-8 (see e.g. claim 1). This overlaps with claim 1.
With respect to claims 2-19, the ‘317 patent claims the same limitations (see e.g. claims 2-19).
With respect to claims 20-30, the ‘317 patent claims the same methods and limitations therein (see e.g. claims 20-30).
3. Claims 1-3, 6, 7, 12-5, 18, and 19 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-3, 5, 9-11, 21, and 29 of U.S. Patent No. 11,505,593 B2. Although the claims at issue are not identical, they are not patentably distinct from each other because the ‘593 patent claims an overlapping composition.
The ‘593 patent claims a prefilled syringe containing aflibercept produced in CDM, wherein the aflibercept contains oxidized variants comprising an oxidized tryptophan and/or histidine residue (see e.g. claim 1). The ‘593 patent further claims 40-250 mg/ml concentrations (see e.g. claim 5). The syringe can also be at pH 5.5-7.5 and contain an excipient (see e.g. claims 10 and 11). This overlaps with claim 1.
With respect to claims 2 and 3, the ‘593 patent as set forth above claims oxidized histidine and/or tryptophan residues (see e.g. claims 1-3).
With respect to claims 6 and 7, as set forth above ‘593 claims an overlapping pH range.
With respect to claims 12-15, the ’593 patent claims oxidized variants at 0.1-1% (see e.g. claims 21 and 29).
With respect to claims 18 and 19, the ‘593 patent claims an autoinjector syringe (see e.g. claims 1 and 9).
Conclusion
No claims are allowed.

APPLICANT REBUTTAL: REMARKS
Claims 1-30 are pending in this application. Claims 27, 29, and 30 are currently amended. No new subject matter is added.
Double Patenting
The Office Action states that pending claims of the Application have been rejected on the ground of non-statutory double patenting as being unpatentable over:
" Claims 1-15, 18-28, and 30 are rejected over claims 13, 17, 20-25, and 28 of U.S. Patent
No. 11,958,894 B2.
" Claims 1-30 over claims 1-30 of U.S. Patent No. 11,542,317 Bl.
" Claims 1-3, 6, 7, 12-15, 18, and 19 over claims 1-3, 5, 9-11, 21, and 29 of U.S. Patent
No. 11,505,593 B2.
In response to the above rejection, Applicant respectfully submits that a terminal disclaimer has been filed in compliance with 37 CFR 1.321(c). As a result, the non-statutory double patenting rejections are overcome and withdrawal is respectfully requested.
In view of the foregoing, Applicant respectfully submits that the present application is in condition for allowance and early notice thereof is earnestly solicited. If any issues remain, the Examiner is requested to call the undersigned at (212) 715-9488.
It is believed that no fees are due in connection with this submission. However, the
Commissioner is hereby authorized to charge any fees due or credit any overpayment to Deposit Account No.
Respectfully submitted,

NEXT OFFICE ACTION: ALLOWANCE
Notice of Pre-AIA  or AIA  Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Application
The entry into the TrackOne program as of 19 November 2024 is acknowledged.
The amendment of 4 February 2025 is entered.
Claims 1-30 are pending and are being examined on the merits.
The Terminal Disclaimer of 4 February 2025 is entered.
The rejections for nonstatutory double patenting are withdrawn in light of the TD filed 4 February 2025.
Allowable Subject Matter
Claims 1-30 are allowed.
The following is an examiner’s statement of reasons for allowance: The prior art does not disclose or suggest a formulation comprising aflibercept produced in a cell culture supplemented with CDM where the aflibercept comprises at least one oxidized amino acid selected from tryptophan, histidine, and combinations thereof. As discussed in the parent 16/996,293 application, US 2021/0010025 discloses oxidation of aflibercept at methionine residues, but does not discuss or contemplate oxidation at histidine or tryptophan residues. Other art, US 2018/0326126 A and WO 2020/229584 A1 explicitly investigate oxidation of aflibercept and never observe tryptophan or histidine oxidation events. The skilled artisan would conclude from this prior art that the aflibercept containing oxidized tryptophan residues, histidine residues, or combinations thereof did not exist nor was it suggested by the prior art. Therefore, the claimed formulation and related methods of utilizing the formulation are novel and unobvious.
Any comments considered necessary by applicant must be submitted no later than the payment of the issue fee and, to avoid processing delays, should preferably accompany the issue fee.  Such submissions should be clearly labeled “Comments on Statement of Reasons for Allowance.”
Conclusion

CITED REFERENCE SUMMARIES: Ref US11958894B2: Claims:
What is claimed is: 1. An oxo-aflibercept composition produced in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein one or more tryptophan amino acid residues of aflibercept is oxidized. 2. The oxo-aflibercept of claim 1, wherein said oxo-aflibercept has a glycosylation profile based on molar percentage selected from the group consisting of about 40% to about 50% total fucosylated glycans, about 30% to about 55% total sialylated glycans, about 2% to about 15% mannose-5, and about 60% to about 79% galactosylated glycans. 3. The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 51.4% to about 52.4% total fucosylated glycans. 4. The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 58.2% to about 59.2% total sialylated glycans. 5. The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 5.19% to about 5.24% mannose-5. 6. The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 79.6% to about 80.6% galactosylated glycans. 7. The oxo-aflibercept composition of claim 1, wherein said oxo-aflibercept is glycosylated at one or more Asparagines as follows: GO-G1cNAc glycosylation; G1-G1cNAc glycosylation; G1S-G1cNAc glycosylation; GO glycosylation; G1 glycosylation; GIS glycosylation; G2 glycosylation; G2S glycosylation; G2S2 glycosylation; GOF glycosylation; G2F2S glycosylation; G2F2S2 glycosylation; G1F glycosylation; G1FS glycosylation; G2F glycosylation; G2FS glycosylation; G2FS2 glycosylation; G3FS glycosylation; G3FS3 glycosylation; GO-2G1cNAc glycosylation; Man4 glycosylation; Man4 A1 G1 glycosylation; Man4 A1 G1S1 glycosylation; ManS glycosylation; Man5 A1 G1 glycosylation; Man5 A1 G1S1 glycosylation; Man6 glycosylation; Man6 GO+Phosphate glycosylation; Man6+Phosphate glycosylation; and/or Man7 glycosylation. 8. An oxo-aflibercept composition produced in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is histidine and/or tryptophan. 9. The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn36 is between 98.3% and 99.4%. 10. The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn68 is between 51.9% and 64.9%. 11. The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn123 is between 98.4% and 99.9%. 12. The oxo-aflibercept composition of claim 11, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn196 is between 55.1% and 98.6%. 13. A prefilled syringe comprising aflibercept produced in a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof. 14. The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 40% to about 50% total fucosylated glycans. 15. The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 30% to about 55% total sialylated glycans. 16. The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 2% to about 15% mannose-5. 17. The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 60% to about 79% galactosylated glycans. 18. The prefilled syringe of claim 16, wherein said aflibercept in said prefilled syringe has a b* value between 0.5 and 5, when the concentration of said aflibercept is normalized to 5.0 g/L. 19. The prefilled syringe of claim 17, wherein said aflibercept in said prefilled syringe has a BY value of at least 6 when the concentration of aflibercept is normalized to 5 g/L protein concentration. 20. The prefilled syringe of claim 16, wherein said aflibercept in said prefilled syringe has between 0.1% and 1% oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof. 21. The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has between 0.2% and 1% oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof. 22. The prefilled syringe of claim 17, wherein said aflibercept loaded into said prefilled syringe is formulated with (a) 40 to 250 mg/ml of aflibercept (b) a pH between 5.5 and 7.5, and (c) one or more pharmaceutically acceptable excipients selected from the group of buffering agents, preservatives, detergents and combinations thereof. 23. The prefilled syringe of claim 17, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer. 24. The prefilled syringe of claim 21, further comprising 0.01% to 5.0% of an organic co-solvent. 25. The prefilled syringe of claim 21, further comprising 30 mM to 150 mM of a tonicity agent. 26. The prefilled syringe of claim 20, wherein 90% of said aflibercept or more by weight is not present as an aggregate. 27. The prefilled syringe of claim 22, wherein said prefilled syringe is glass. 28. The prefilled syringe of claim 22, wherein said prefilled syringe is an auto injector. 29. The prefilled syringe of claim 13, wherein said aflibercept is glycosylated at one or more Asparagines as follows: GO-G1cNAc glycosylation; G1-G1cNAc glycosylation; G1S-G1cNAc glycosylation; GO glycosylation; G1 glycosylation; G15 glycosylation; G2 glycosylation; G2S glycosylation; G2S2 glycosylation; GOF glycosylation; G2F2S glycosylation; G2F2S2 glycosylation; G1F glycosylation; G1FS glycosylation; G2F glycosylation; G2FS glycosylation; G2FS2 glycosylation; G3FS glycosylation; G3FS3 glycosylation; GO-2G1cNAc glycosylation; Man4 glycosylation; Man4 A1G1 glycosylation; Man4 A1 G151 glycosylation; ManS glycosylation; Man5 A1 G1 glycosylation; Man5 A1 G151 glycosylation; Man6 glycosylation; Man6 GO+Phosphate glycosylation; Man6+Phosphate glycosylation; and/or Man7 glycosylation. 30. The prefilled syringe of claim 13, wherein said aflibercept has a glycosylation profile based on molar percentage selected from the group consisting of about 40% to about 50% total fucosylated glycans, about 30% to about 55% total sialylated glycans, about 2% to about 15% mannose-5, and about 60% to about 79% galactosylated glycans.
An oxo-aflibercept composition produced in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein one or more tryptophan amino acid residues of aflibercept is oxidized.
The oxo-aflibercept of claim 1, wherein said oxo-aflibercept has a glycosylation profile based on molar percentage selected from the group consisting of about 40% to about 50% total fucosylated glycans, about 30% to about 55% total sialylated glycans, about 2% to about 15% mannose-5, and about 60% to about 79% galactosylated glycans.
The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 51.4% to about 52.4% total fucosylated glycans.
The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 58.2% to about 59.2% total sialylated glycans.
The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 5.19% to about 5.24% mannose-5.
The oxo-aflibercept composition of claim 2, wherein said oxo-aflibercept has about 79.6% to about 80.6% galactosylated glycans.
The oxo-aflibercept composition of claim 1, wherein said oxo-aflibercept is glycosylated at one or more Asparagines as follows: GO-G1cNAc glycosylation; G1-G1cNAc glycosylation; G1S-G1cNAc glycosylation; GO glycosylation; G1 glycosylation; GIS glycosylation; G2 glycosylation; G2S glycosylation; G2S2 glycosylation; GOF glycosylation; G2F2S glycosylation; G2F2S2 glycosylation; G1F glycosylation; G1FS glycosylation; G2F glycosylation; G2FS glycosylation; G2FS2 glycosylation; G3FS glycosylation; G3FS3 glycosylation; GO-2G1cNAc glycosylation; Man4 glycosylation; Man4 A1 G1 glycosylation; Man4 A1 G1S1 glycosylation; ManS glycosylation; Man5 A1 G1 glycosylation; Man5 A1 G1S1 glycosylation; Man6 glycosylation; Man6 GO+Phosphate glycosylation; Man6+Phosphate glycosylation; and/or Man7 glycosylation.
An oxo-aflibercept composition produced in a cell culture media comprising a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is histidine and/or tryptophan.
The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn36 is between 98.3% and 99.4%.
The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn68 is between 51.9% and 64.9%.
The oxo-aflibercept composition of claim 8, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn123 is between 98.4% and 99.9%.
The oxo-aflibercept composition of claim 11, wherein N-glycosylation of the F(ab′)2 portion of said oxo-aflibercept at Asn196 is between 55.1% and 98.6%.
A prefilled syringe comprising aflibercept produced in a cell culture media wherein chemically defined media (CDM) is added to cell culture media and wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof.
The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 40% to about 50% total fucosylated glycans.
The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 30% to about 55% total sialylated glycans.
The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 2% to about 15% mannose-5.
The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has about 60% to about 79% galactosylated glycans.
The prefilled syringe of claim 16, wherein said aflibercept in said prefilled syringe has a b* value between 0.5 and 5, when the concentration of said aflibercept is normalized to 5.0 g/L.
The prefilled syringe of claim 17, wherein said aflibercept in said prefilled syringe has a BY value of at least 6 when the concentration of aflibercept is normalized to 5 g/L protein concentration.
The prefilled syringe of claim 16, wherein said aflibercept in said prefilled syringe has between 0.1% and 1% oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof.
The prefilled syringe of claim 13, wherein said aflibercept in said prefilled syringe has between 0.2% and 1% oxidized amino acid residue selected from the group consisting of His86, His110, His145, His209, His95, His19, His203, Trp58, Trp138 and a combination thereof.
The prefilled syringe of claim 17, wherein said aflibercept loaded into said prefilled syringe is formulated with (a) 40 to 250 mg/ml of aflibercept (b) a pH between 5.5 and 7.5, and (c) one or more pharmaceutically acceptable excipients selected from the group of buffering agents, preservatives, detergents and combinations thereof.
The prefilled syringe of claim 17, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer.
The prefilled syringe of claim 21, further comprising 0.01% to 5.0% of an organic co-solvent.
The prefilled syringe of claim 21, further comprising 30 mM to 150 mM of a tonicity agent.
The prefilled syringe of claim 20, wherein 90% of said aflibercept or more by weight is not present as an aggregate.
The prefilled syringe of claim 22, wherein said prefilled syringe is glass.
The prefilled syringe of claim 22, wherein said prefilled syringe is an auto injector.
The prefilled syringe of claim 13, wherein said aflibercept is glycosylated at one or more Asparagines as follows: GO-G1cNAc glycosylation; G1-G1cNAc glycosylation; G1S-G1cNAc glycosylation; GO glycosylation; G1 glycosylation; G15 glycosylation; G2 glycosylation; G2S glycosylation; G2S2 glycosylation; GOF glycosylation; G2F2S glycosylation; G2F2S2 glycosylation; G1F glycosylation; G1FS glycosylation; G2F glycosylation; G2FS glycosylation; G2FS2 glycosylation; G3FS glycosylation; G3FS3 glycosylation; GO-2G1cNAc glycosylation; Man4 glycosylation; Man4 A1G1 glycosylation; Man4 A1 G151 glycosylation; ManS glycosylation; Man5 A1 G1 glycosylation; Man5 A1 G151 glycosylation; Man6 glycosylation; Man6 GO+Phosphate glycosylation; Man6+Phosphate glycosylation; and/or Man7 glycosylation.
The prefilled syringe of claim 13, wherein said aflibercept has a glycosylation profile based on molar percentage selected from the group consisting of about 40% to about 50% total fucosylated glycans, about 30% to about 55% total sialylated glycans, about 2% to about 15% mannose-5, and about 60% to about 79% galactosylated glycans.
Ref US11542317B1: Claims:
What is claimed is: 1. A formulation comprising: (a) 40 to 200 mg/ml of aflibercept produced in chemically defined media (CDM), wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof; (b) one or more pharmaceutically acceptable excipients selected from the group of buffering agents, preservatives, detergents and combinations thereof; and (d) said formulation having a pH between 5.0 and 8.0. 2. The formulation of claim 1, wherein said oxidized amino acid residue is histidine. 3. The formulation of claim 1, wherein said oxidized amino acid residue is tryptophan. 4. The formulation of claim 2, wherein said one or more excipients includes 5-40 mM of a buffer. 5. The formulation of claim 3, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer. 6. The formulation of claim 2, wherein the pH is between 5.5 and 7.0. 7. The formulation of claim 3, wherein the pH is between 5.5 and 7.0. 8. The formulation of claim 2, further comprising 0.01% to 5.0% of an organic co-solvent. 9. The formulation of claim 3, further comprising 0.01% to 5.0% of an organic co-solvent. 10. The formulation of claim 2, further comprising 30 mM to 150 mM of a tonicity agent. 11. The formulation of claim 3, further comprising 30 mM to 150 mM of a tonicity agent. 12. The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise less than 1.0% of the total amount of aflibercept. 13. The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise between 0.05% and 1.0% of the total amount of aflibercept. 14. The formulation of claim 3, wherein said oxidized tryptophan variants of aflibercept comprise less than 1.0% of the total amount of aflibercept. 15. The formulation of claim 3, wherein said oxidized tryptophan variants of aflibercept comprise between 0.2% and 1.0% of the total amount of aflibercept. 16. The formulation of claim 1, wherein said formulation is stored under an inert gas. 17. The formulation of claim 1, wherein said formulation is stored in a vial. 18. The formulation of claim 1, wherein said formulation is stored in a syringe. 19. The formulation of claim 18, wherein said formulation in said syringe is an autoinjector syringe. 20. A method of administering a pharmaceutically effective amount of aflibercept by intravitreal injection wherein said aflibercept is made in chemically defined media (CDM) and includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof and one or more pharmaceutically acceptable excipients. 21. The method of claim 20, wherein the pharmaceutically effective amount of aflibercept is 2 mg. 22. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection once every 4 weeks. 23. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection once every 4 weeks for the first 3 months, followed by 2 mg via intravitreal injection once every 8 weeks. 24. The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection every 4 weeks for the first 5 injections followed by 2 mg via intravitreal injection once every 8 weeks. 25. The method of claim 20, further comprising the step of administering an anesthesia, an antiseptic and/or an antibiotic. 26. A method of treating Neovascular (Wet) Age-Related Macular Degeneration (AMD), Macular Edema Following Retinal Vein Occlusion (RVO), Diabetic Macular Edema (DME), or Diabetic Retinopathy with aflibercept wherein said aflibercept is made in chemically defined media (CDM) and includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof. 27. The method claim 26, further comprising a reduction in mean retinal thickness. 28. The method claim 26, further comprising an increase in visual acuity. 29. The method of claim 26, further comprising a ≥2 step improvement on an Early Treatment of Diabetic Retinopathy Scale—Diabetic Retinopathy Severity Scale (ETDRS-DRSS) from baseline. 30. The method claim 26, further comprising a stabilization or elimination of one or more symptoms or indicia of Neovascular (Wet) Age-Related Macular Degeneration (AMD), Macular Edema Following Retinal Vein Occlusion (RVO), Diabetic Macular Edema (DME), or Diabetic Retinopathy by any clinically measurable degree.
A formulation comprising: (a) 40 to 200 mg/ml of aflibercept produced in chemically defined media (CDM), wherein said aflibercept includes oxidized variants that have at least one oxidized amino acid residue selected from the group consisting of tryptophan, histidine, and a combination thereof; (b) one or more pharmaceutically acceptable excipients selected from the group of buffering agents, preservatives, detergents and combinations thereof; and (d) said formulation having a pH between 5.0 and 8.0.
The formulation of claim 1, wherein said oxidized amino acid residue is histidine.
The formulation of claim 1, wherein said oxidized amino acid residue is tryptophan.
The formulation of claim 2, wherein said one or more excipients includes 5-40 mM of a buffer.
The formulation of claim 3, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer.
The formulation of claim 2, wherein the pH is between 5.5 and 7.0.
The formulation of claim 3, wherein the pH is between 5.5 and 7.0.
The formulation of claim 2, further comprising 0.01% to 5.0% of an organic co-solvent.
The formulation of claim 3, further comprising 0.01% to 5.0% of an organic co-solvent.
The formulation of claim 2, further comprising 30 mM to 150 mM of a tonicity agent.
The formulation of claim 3, further comprising 30 mM to 150 mM of a tonicity agent.
The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise less than 1.0% of the total amount of aflibercept.
The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise between 0.05% and 1.0% of the total amoun
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