Anti-VEGF protein compositions and methods for producing the same

Patent metadata

Field	Value
Publication number	US-12319728-B2
Application number	US-202418637124-A
Country	US
Kind code	B2
Filing date	Apr 16, 2024
Priority date	Dec 6, 2019
Publication date	Jun 3, 2025
Grant date	Jun 3, 2025

Abstract

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.

First claim

What is claimed is:

1. A composition comprising oxo-aflibercept, wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is phenylalanine and/or tyrosine.

2. The composition 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 composition of claim 1 , further comprising a pharmaceutically acceptable carrier.

4. The composition of claim 3 , wherein said oxo-aflibercept is freeze-dried.

5. The composition of claim 4 , wherein said composition is sterile.

6. The composition of claim 1 , wherein said oxo-aflibercept is enzymatically digested resulting in one or more oligopeptides, and wherein said one or more oligopeptides is selected from the group consisting of: SEQ ID NO. 69, SEQ ID NO. 70, SEQ ID NO. 32, and combinations thereof.

7. The composition of claim 6 , wherein said enzymatic digestion is performed using trypsin.

8. An oxo-aflibercept composition produced in a cell culture media, wherein CDM is added to said cell culture media, wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is phenylalanine and/or tyrosine.

9. The oxo-aflibercept composition of claim 8 , wherein said oxo-aflibercept composition 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.

10. The oxo-aflibercept composition of claim 9 , wherein soy hydrolysate media is added to said cell culture media.

11. The oxo-aflibercept composition of claim 8 , wherein components derived from soy hydrolysate are added to said cell culture media.

12. The oxo-aflibercept composition of claim 8 , wherein plant or animal-derived components are added to said cell culture media.

13. The composition of claim 8 , wherein said oxo-aflibercept is enzymatically digested resulting in one or more oligopeptides, and wherein said one or more oligopeptides is selected from the group consisting of: SEQ ID NO. 69, SEQ ID NO. 70, SEQ ID NO. 32, and combinations thereof.

14. The composition of claim 13 , wherein said enzymatic digestion is performed using trypsin.

15. An oxo-aflibercept composition produced in a cell culture media, wherein CDM is a majority component of said cell culture media, 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.

16. The oxo-aflibercept composition of claim 15 , wherein said oxo-aflibercept composition 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.

17. The oxo-aflibercept composition of claim 16 , wherein said cell culture media is supplemented with soy hydrolysate media.

18. The oxo-aflibercept composition of claim 15 , wherein components derived from soy hydrolysate are added to said cell culture media.

19. The oxo-aflibercept composition of claim 15 , wherein plant or animal-derived components are added to cell culture media.

20. The composition of claim 15 , wherein said oxo-aflibercept is enzymatically digested resulting in one or more oligopeptides, and wherein said one or more oligopeptides is selected from the group consisting of: SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21, SEQ ID NO. 22, SEQ ID NO. 23, SEQ ID NO. 28, SEQ ID NO. 29, SEQ ID NO. 30, SEQ ID NO. 31, SEQ ID NO. 32 and combinations thereof.

21. The composition of claim 20 , wherein said enzymatic digestion is performed using trypsin.

22. The composition of claim 15 , further comprising a pharmaceutically acceptable carrier.

23. An oxo-aflibercept composition produced in a cell culture media, wherein CDM is a majority component of said cell culture media, wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is phenylalanine and/or tyrosine.

24. The oxo-aflibercept composition of claim 23 , wherein said oxo-aflibercept composition 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.

25. The oxo-aflibercept composition of claim 23 , wherein said cell culture media is supplemented with soy hydrolysate media.

26. The oxo-aflibercept composition of claim 23 , wherein components derived from soy hydrolysate are added to said cell culture media.

27. The oxo-aflibercept composition of claim 23 , wherein plant or animal-derived components are added to cell culture media.

28. The composition of claim 23 , wherein said oxo-aflibercept is enzymatically digested resulting in one or more oligopeptides, and wherein said one or more oligopeptides is selected from the group consisting of: SEQ ID NO. 69, SEQ ID NO. 70, SEQ ID NO. 32 and combinations thereof.

29. The composition of claim 28 , wherein said enzymatic digestion is performed using trypsin.

30. The composition of claim 23 , further comprising a pharmaceutically acceptable carrier.

Assignees

• Regeneron Pharma

Inventors

• Wang Shunhai
• Li Ning
• Chen Hunter
• Bhupender Bhalla Amardeep Singh
• Lawrence Shawn M
• Johnson Amy S
• Casey Meghan
• Grapel Jaimie

Classifications

• C12N2529/10

  Stimulation by light · CPC title

• C12N15/85

  for animal cells · CPC title

• C07K1/06

  using protecting groups or activating agents {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C12N2510/02

  Cells for production · CPC title

• C07K1/165

  mixed-mode chromatography · CPC title