Office Action 2: Notice of Allowance (2025-02-24)

CLAIMS:
Listing of Claims:

The following claim amendments will replace all prior versions in this application.

What is claimed is:

1\. (Original) 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\. (Original) The formulation of claim 1, wherein said oxidized amino acid residue is histidine.

3\. (Original) The formulation of claim 1, wherein said oxidized amino acid residue is tryptophan.

4\. (Original) The formulation of claim 2, wherein said one or more excipients includes 5-40 mM of a buffer.

5\. (Original) The formulation of claim 3, wherein said one or more pharmaceutically acceptable excipients includes 5-40 mM of a buffer.

6\. (Original) The formulation of claim 2, wherein the pH is between 5.5 and 7.0.

7\. (Original) The formulation of claim 3, wherein the pH is between 5.5 and 7.0.

8\. (Original) The formulation of claim 2, further comprising 0.01% to 5.0% of an organic co- solvent.

9\. (Original) The formulation of claim 3, further comprising 0.01% to 5.0% of an organic co- solvent.

10\. (Original) The formulation of claim 2, further comprising 30 mM to 150 mM of a tonicity agent.

11\. (Original) The formulation of claim 3, further comprising 30 mM to 150 mM of a tonicity agent.

12\. (Original) The formulation of claim 2, wherein said oxidized histidine variants of aflibercept comprise less than 1.0% of the total amount of aflibercept.

13\. (Original) 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\. (Original) The formulation of claim 3, wherein said oxidized tryptophan variants of aflibercept comprise less than 1.0% of the total amount of aflibercept.

15\. (Original 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\. (Original) The formulation of claim 1, wherein said formulation is stored under an inert gas.

17\. (Original) The formulation of claim 1, wherein said formulation is stored in a vial.

18\. (Original) The formulation of claim 1, wherein said formulation is stored in a syringe.

19\. (Original) The formulation of claim 18, wherein said formulation in said syringe is an autoinjector syringe.

20\. (Original) 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\. (Original) The method of claim 20, wherein the pharmaceutically effective amount of aflibercept is 2 mg.

22\. (Original) The method of claim 21, wherein the pharmaceutically effective amount of aflibercept is administered by intravitreal injection once every 4 weeks.

23\. (Original) 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\. (Original) 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\. (Original) The method of claim 20, further comprising the step of administering an anesthesia, an antiseptic and/or an antibiotic.

26\. (Original) 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\. (Currently Amended) The method ofclaim 26, further comprising a reduction in mean retinal thickness.

28\. (Currently Amended) The method ofclaim 26, further comprising an increase in visual acuity.

29\. (Original) 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\. (Currently Amended) The method ofclaim 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:
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

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,


CITED REFERENCE SUMMARIES: Ref US20210010025A1: [content unavailable]
Ref US20180326126A1: Claims:
A pre-filled syringe containing a liquid formulation of a VEGF antagonist and comprising a syringe barrel, wherein the syringe barrel is made of plastic and is silicone-free, and further comprising a non-retractable stopper.
A pre-filled syringe containing a liquid formulation of a VEGF antagonist and comprising a syringe barrel, wherein the syringe barrel is made of plastic, is silicone-free, and has a length of 45 mm to 65 mm.
The pre-filled syringe according to claim 1 , wherein the VEGF antagonist is an anti-VEGF antibody or an antigen-binding fragment of such antibody or a VEGF receptor fusion protein.
The pre-filled syringe according to claim 3 , anti-VEGF antagonist is ranibizumab or aflibercept.
The pre-filled syringe according to claim 1 , wherein the antagonist concentration is 1 to 100 mg/ml.
The pre-filled syringe according to claim 1 , containing less than 50 particles per ml of the liquid formulation having a diameter of 10 Î¼m or greater.
The pre-filled syringe according to claim 1 , containing less than 5 particles per ml of the liquid formulation having a diameter of 25 Î¼m or greater.
The pre-filled syringe according to claim 1 , having a sliding force of less than or equal to 10N.
The pre-filled syringe according to claim 1 , further comprising a silicone-free stopper.
The pre-filled syringe according to claim 1 , wherein the syringe barrel is made of cycloolefin polymer or cycloolefin copolymer.
The pre-filled syringe according to claim 1 , wherein the syringe barrel comprises an internal coating other than a silicone coating.
The pre-filled syringe according to claim 1 , comprising a staked needle.
A kit comprising one or more pre-filled syringes of claim 1 .
The pre-filled syringe according to claim 1 for use in administering a liquid formulation of a VEGF antagonist to a patient having an ocular disease.
The pre-filled syringe for the use according to claim 13 , wherein the ocular disease is selected from the group consisting of age-related macular degeneration (AMD), visual impairment due to diabetic macular oedema (DME), visual impairment due to macular oedema secondary to retinal vein occlusion (branch RVO or central RVO), diabetic retinopathy in patients with diabetic macular edema, or visual impairment due to choroidal neovascularisation (CNV) secondary to pathologic myopia.
The pre-filled syringe for the use according to claim 14 or 15 , wherein a volume of 30 to 100 Î¼l of the liquid formulation is administered to the patient.
Ref WO2020229584A1: Claims:
Method for reducing the percentage of oxidized methionine residues in a recombinant protein, said method comprising culturing cells expressing said recombinant protein in a suitable cell culture medium and feeding the cells with glutamine.
Method according to claim 1, comprising feeding the cells with 0.2 mM to 5 mM glutamine.
Method according to claim 1 or 2, comprising feeding the cells with 0.5 mM to 4 mM glutamine on day 3 to 5 of the culture.
Method according to any one of the preceding claims, comprising feeding the cells with 0.2 mM to 2 mM glutamine on day 6 to 12 of the culture.
Method according to any one of the preceding claims, further comprising feeding the cells with methionine.
Method according to claim 5, comprising feeding the cells with 250 to 1000 mg/1 methionine.
Method according to any one of the preceding claims, further comprising reducing the culture temperature.
Method according to claim 7, wherein the culture temperature is reduced from 37Â°C to a temperature within the range of 30Â°C to 35Â°C.
Method according to claim 7 or 8, wherein the temperature is reduced in a first step from a first temperature of 37Â°C to a second temperature within the range of 33Â°C to 35Â°C and in a second step from the second temperature of 33Â°C to 35Â°C to a third temperature within the range of 31Â°C to 33Â°C.
Method according to any one of the preceding claims, wherein the recombinant protein is a recombinant antibody or a fusion protein of a receptor and the Fc portion of an immunoglobulin.
Method according to any one of the preceding claims, wherein the recombinant protein is aflibercept.
Method according to any one of the preceding claims, wherein the cells are CHO cells.
Use of glutamine for reducing the percentage of oxidized methionine residues in a composition comprising a recombinant protein.
Method for producing a recombinant protein, comprising
(a) culturing cells expressing the recombinant protein in a suitable cell culture medium;
(b) feeding the cells with glutamine, wherein the cells are fed with 0.5 mM to 4 mM glutamine on day 3 to 5 of the culture and with 0.2 mM to 2 mM glutamine on day 6 to 12 of the culture; and
(c) obtaining the recombinant protein.
Method for producing aflibercept, comprising:
(a) culturing cells expressing aflibercept in a suitable cell culture medium;
(b) feeding the cells with glutamine; and
(c) obtaining the aflibercept.
Ref US20210010025: Claims:
1. An expression construct comprising an expression cassette flanked by AAV inverted terminal repeats (ITRs) wherein the expression cassette comprises a transgene encoding a VEGF-TrapHuPTM operably linked to one or more regulatory sequences that control expression of the transgene in human retinal cells or human liver cells, wherein the transgene encodes a leader sequence operable in human retinal cells or human liver cells and a VEGF-TrapHuPTM, wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 204 of SEQ ID NO: 1.
2. The expression construct of claim 1 wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 205 of SEQ ID NO: 1 linked at the C terminus to an IgG1, IgG2, or IgG4 Fc region comprising at least a partial hinge region at the N-terminus of the Fc region.
3. The expression construct of claim 2, wherein the Fc region comprises a full hinge region.
4. The expression construct of claim 2, wherein one or more of the cysteine residues within the hinge region is substituted with a serine.
5. The expression construct of claim 2, wherein the Fc region has one or more amino acid substitutions which reduce FcRn binding compared to the Fc region without the amino acid substitutions.
6. The expression construct of claim 1 wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 205 of SEQ ID NO: 1 linked at the C terminus to an Ig-like domain of Flt-1 or KDR.
7. The expression construct of claim 1, wherein the expression construct comprises a second VEGF-TrapHuPTM comprising an amino acid sequence having amino acid residues 1 to 204 of SEQ ID NO: 1.
8. The expression construct of claim 1 wherein the VEGF-TrapHuPTM has an amino acid sequence selected from
i. the amino acid sequence of SEQ ID NO: 1 (FIG. 1),
ii. the amino acid sequence of SEQ ID NO: 1 with an alanine substitution at position 238 and/or 295 and/or an alanine or glutamine substitution at position 420;
iii. the amino acid sequence of SEQ ID NO: 1 with an alanine or glutamine substitution at position 420 (FIG. 3);
iv. the amino acid sequence of amino acid residues 1 to 205 of SEQ ID NO: 1 and optionally linked to the C-terminus a sequence selected from SEQ ID Nos: 46 to 48 (FIG. 4);
v. the amino acid sequence consisting of residues 1 to 204 of SEQ ID NO: 1;
vi. the amino acid sequence of amino acid sequence residues 1 to 205 of SEQ ID NO: 1 linked at the C terminus to one of the amino acid sequences of SEQ ID NOs: 19, 20, 49, 50, 51, 52, 53, or 54 (FIG. 7C-7H); and
vii. the amino acid sequence of amino acid sequence residues 1 to 205 of SEQ ID NO: 1 linked at the C terminus to either SEQ ID NO: 55 or 56. (FIG. 8C/8D)
9. The expression construct of clam 1, wherein the leader sequence is one of SEQ ID Nos:
36 to 39 or 59 to 67. (retinal cells)
10. The expression construct of claim 1, wherein the leader sequence is one of SEQ ID Nos: 68 to 87 or 97. (liver cells)
11. The expression construct of claim 1, wherein at least one of the regulatory sequences is a constitutive promoter.
12. The expression construct of claim 1, wherein the one or more regulatory sequences are a CB7 promoter, a chicken β-actin intron and a rabbit β-globin poly A signal.
13. The expression construct of claim 1, wherein at least one of the regulatory sequences is an inducible promoter, optionally a hypoxia-inducible promoter or a rapamycin inducible promoter.
14. An adeno-associated virus (AAV) vector comprising a viral capsid that is at least 95% identical to the amino acid sequence of an AAV8 capsid (SEQ ID NO: 11) or AAV2 capsid (SEQ ID NO: 5) or is a variant of AAV8 or AAV2, and a viral genome comprising an expression construct of claim 1.
15. The AAV vector of claim 14, wherein the viral capsid is AAV.7m8.
16. A pharmaceutical composition for ocular administration comprising an AAV vector comprising:
a viral capsid that is at least 95% identical to the amino acid sequence of an AAV8 capsid (SEQ ID NO: 11) or AAV2 capsid (SEQ ID NO: 5) or is a variant of AAV8 or AAV2; and a viral genome comprising an expression construct of claim 1;
wherein said AAV vector is formulated for subretinal, intravitreal or suprachororidal administration to the eye of said subject.
17. The pharmaceutical composition of claim 16, wherein the viral capsid is AAV.7m8.
18. A pharmaceutical composition for intravenous administration comprising an AAV vector comprising:
a viral capsid that is at least 95% identical to the amino acid sequence of an AAV8 capsid (SEQ ID NO: 11) or is a variant of AAV8; and a viral genome comprising an expression construct of claim 1;
wherein said AAV vector is formulated for intravenous administration to said subject.
19. A method of treating a human subject diagnosed with metastatic colon cancer or an eye related disorder selected from neovascular age-related macular degeneration (nAMD), diabetic retinopathy, diabetic macular edema (DME), central retinal vein occlusion (RVO), pathologic myopia, or polypoidal choroidal vasculopathy, said method comprising delivering to the retina of said human subject with the eye-related disorder or to the cancer cells or neovascularized tissue around said cancer cells of said human subject with metastatic colon cancer, a therapeutically effective amount of VEGF-TrapHuPTM produced by human liver cells or human retinal cells selected from human photoreceptor cells (cone cells, rod cells); horizontal cells; bipolar cells; amacrine cells; retina ganglion cells (midget cell, parasol cell, bistratified cell, giant retina ganglion cell, photosensitive ganglion cell, and mullerglia); and retinal pigment epithelial cells, wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 204 of SEQ ID NO: 1.
20. A method of treating a human subject diagnosed metastatic colon cancer or an eye related disorder selected from neovascular age-related macular degeneration (nAMD), diabetic retinopathy, diabetic macular edema (DME), central retinal vein occlusion (RVO), pathologic myopia, or polypoidal choroidal vasculopathy, said method comprising delivering to the retina of said human subject with the eye-related disorder or to the cancer cells or neovascularized tissue around said cancer cells of said human subject with metastatic colon cancer, a therapeutically effective amount of a VEGF-TrapHuPTM containing an α2,6-sialylated glycan and/or a tyrosine sulfation, wherein the VEGF-TrapHuPTM comprises an amino acid sequence having amino acid residues 1 to 204 of SEQ ID NO: 1.
21. The method of claim 20, wherein the VEGF-TrapHuPTM expressed does not contain detectable NeuGc or α-Gal.
22. A method of treating a human subject diagnosed with metastatic colon cancer or an eye related disorder selected from neovascular age-related macular degeneration (nAMD), diabetic retinopathy, diabetic macular edema (DME), central retinal vein occlusion (RVO), pathologic myopia, or polypoidal choroidal vasculopathy, said method comprising: administering to the liver of said human subject with metastatic colon cancer and to the the subretinal space in the eye of said human subject with the eye-related disorder, a therapeutically effective amount of a recombinant nucleotide expression vector comprising an expression construct of claims 1, wherein VEGF-TrapHuPTM expressed in the liver contains a α2,6-sialylated glycan or tyrosine-sulfation.
23. The method of claim 22, wherein the VEGF-TrapHuPTM expressed does not contain detectable NeuGc or α-Gal.
24. The method of claim 22, wherein the recombinant nucleotide expression vector is an AAV8 viral vector or an AAV2 viral vector or an AAV viral vector that is a variant of AVV2 or AAV8.
25. The method of claim 24, wherein the recombinant nucleotide expression vector is an AAV.7m8 viral vector.
26. A method of manufacturing an AAV2 or AAV8 viral vector comprising a VEGF-Trap transgene, said method comprising culturing host cells under conditions appropriate for production of the AAV2 or AAV8 viral vector, wherein the host cells are stably transformed with a nucleic acid vector comprise an expression construct of claim 1 comprising nucleotide sequences encoding the AAV2 or AAV8 replication and capsid proteins or variants thereof; and recovering the AAV2 or AAV8 viral vector produced by the host cell.
27. The method of claim 26, wherein the viral vector comprises nucleotide sequences encoding the AAV.7m8 replication and capsid proteins.
28. A method of producing recombinant AAVs comprising:
(a) culturing a host cell containing:
(i) an artificial genome comprising an expression construct of claim 1;
(ii) a trans expression cassette lacking AAV ITRs, wherein the trans expression cassette encodes an AAV rep and capsid protein operably linked to expression control elements that drive expression of the AAV rep and capsid proteins in the host cell in culture and supply the rep and cap proteins in trans;
(iii) sufficient adenovirus helper functions to permit replication and packaging of the artificial genome by the AAV capsid proteins; and
 (b) recovering recombinant AAV encapsidating the artificial genome from the cell culture.
Ref US20180326126A: [content unavailable]