Correctly folded etanercept in high purity and excellent yield

• US 10,954,294 B2
• Filed: 06/17/2019
• Issued: 03/23/2021
• Est. Priority Date: 09/11/2012
• Status: Active Grant

• Current Assignee: Coherus BioSciences Inc.
• Original Assignee: Coherus BioSciences Inc.
• Inventors: Arakawa, Tsutomu, Farrar, Douglas
• Primary Examiner(s): D' Ambrosio, Thea
• Application Number:
  US16/443,594
• Publication Number:
  US 20190300601A1
• Time in Patent Office: 645 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 1/165 mixed-mode chromatography
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

A mixed mode chromatography method for separating correctly folded from incorrectly folded conformations of a given protein is provided. The method is highly effective in separating correctly folded etanercept from incorrectly folded etanercept and aggregates in commercially attractive yields capable of affording etanercept preparations having very high purity in terms of correctly folded etanercept versus incorrectly folded etanercept. The invention is further directed to protein preparations and formulations comprising correctly folded proteins obtained using the present methods, and methods of treatment using the high purity preparations obtained from the mixed mode method.

Claims

1. A stable aqueous etanercept-containing pharmaceutical composition comprising:

a) an etanercept-containing protein mixture comprising correctly folded etanercept in an amount constituting greater than 95 wt % of the protein mixture and less than 5 wt % incorrectly folded etanercept, wherein said incorrectly folded etanercept is an etanercept protein that has a conformation different from that of the correctly folded etanercept and said different conformation renders said incorrectly folded etanercept protein lacking in biological activity as a TNF inhibitor and said incorrectly folded etanercept is not an aggregate, wherein said etanercept-containing protein mixture is obtained by a mixed-mode chromatography procedure comprising binding correctly folded etanercept and incorrectly folded etanercept to a mixed-mode chromatography resin having both ion exchange and hydrophobic moieties and contacting the mixed-mode chromatography resin to elute with a salt solution at a pH between 4.5 and 8.5;

b) about 0.1 to 2 weight percent of a correctly folded etanercept-stabilizing combination of sucrose, sodium citrate, and one of alanine, glycine, and lysine;

c) 0 to about 25 mM sodium chloride;

d) a buffer selected from phosphate, histidine, citrate, maleate, tartrate, acetate, tris-(hydroxymethyl)-aminomethane (tris), and bicarbonate, or a combination thereof,

wherein the composition does not contain cysteine and does not contain arginine,

wherein the composition is an aqueous pharmaceutical composition having a pH of about 6.2 to 7.4.

2. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the correctly folded etanercept-stabilizing combination contains sucrose, sodium citrate, and alanine.

3. The stable aqueous etanercept-containing pharmaceutical composition of claim 2, comprising about 25 mM sodium chloride.

4. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the correctly folded etanercept-stabilizing combination contains sucrose, sodium citrate, and glycine.

5. The stable aqueous etanercept-containing pharmaceutical composition of claim 4, comprising about mM sodium chloride.

6. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the correctly folded etanercept-stabilizing combination contains sucrose, sodium citrate, and lysine.

7. The stable aqueous etanercept-containing pharmaceutical composition of claim 6, comprising about 25 mM sodium chloride.

8. The stable aqueous etanercept-containing pharmaceutical composition of claim 7, wherein the composition does not contain any amino acid other than lysine.

9. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein less than 3 wt % of the etanercept in the etanercept-containing protein mixture is incorrectly folded etanercept.

10. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein less than 1 wt % of the etanercept in the etanercept-containing protein mixture is incorrectly folded etanercept.

11. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein at least 98 wt % of the etanercept in the etanercept-containing protein mixture is correctly folded etanercept.

12. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the aqueous buffer is citrate buffer.

13. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the stable aqueous etanercept-containing pharmaceutical composition comprises less than 3 wt % aggregates of correctly folded etanercept.

14. A vial, syringe, or injector pen containing the stable aqueous etanercept-containing pharmaceutical composition of claim 8.

15. A vial, syringe, or injector pen containing the stable aqueous etanercept-containing pharmaceutical composition of claim 12.

16. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 1.

17. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 10.

18. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 11.

19. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 3.

20. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 5.

21. A method of treating a tumor necrosis factor (TNF) mediated disease in a patient comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition according to claim 12.