CORRECTLY FOLDED ETANERCEPT IN HIGH PURITY AND EXCELLENT YIELD

• US 20190300601A1
• Filed: 06/17/2019
• Published: 10/03/2019
• Est. Priority Date: 09/11/2012
• Status: Active Grant

• Current Assignee: Coherus BioSciences Inc.
• Original Assignee: Coherus BioSciences Inc.
• Inventors: ARAKAWA, Tsutomu, FARRAR, Douglas
• Granted Patent: US 10,954,294 B2
• Time in Patent Office: Days
• 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 about 90 wt. % of the protein mixture;

b. about 0.1 to 2 weight percent of one or more excipients selected from sucrose, mannitol, alanine, glycine, and lysine;

c. one or more tonicity modifiers selected from sodium chloride, potassium chloride, sodium sulfate, and sodium citrate;

d. a buffer selected from phosphate, histidine, citrate, maleate, tartrate, succinate, 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 composition is less immunogenic than a commercially available etanercept-containing composition.

3. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the composition contains less incorrectly folded etanercept than a commercially available etanercept-containing composition.

4. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, comprising about 0.1 to about 2 wt. % sucrose;

about 0.1 to 2 wt. % mannitol, alanine, glycine, or lysine; and

about 10 mM to about 200 mM of one or more tonicity modifiers selected from sodium chloride, potassium chloride, sodium sulfate, and sodium citrate.

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

6. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein less than 5 wt. % of the etanercept in the etanercept-containing protein mixture is incorrectly folded etanercept.

7. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein at least 90 wt. % of the etanercept in the etanercept-containing protein mixture is correctly folded etanercept.

8. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, wherein the one or more excipients are lysine and sucrose and the one or more tonicity modifiers are sodium citrate and sodium chloride.

9. The stable aqueous etanercept-containing pharmaceutical composition of claim 8, comprising about 0.1 to about 2 wt. % of the sucrose and lysine and about 10 mM to about 200 mM of the sodium citrate and the sodium chloride.

10. The stable aqueous etanercept-containing pharmaceutical composition of claim 1, comprising about 0.1 to about 2 wt. % of a combination of sucrose, sodium chloride, and lysine.

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

12. The stable aqueous etanercept-containing pharmaceutical composition of claim 9, wherein the aqueous buffer is citrate buffer.

13. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 1.

14. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 6.

15. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 7.

16. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 8.

17. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 9.

18. A method of treating a patient in need of treatment with etanercept, comprising administering to said patient the stable aqueous etanercept-containing pharmaceutical composition of claim 12.

19. A vial, syringe, or injector pen containing the stable aqueous etanercept-containing pharmaceutical composition of claim 9.

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