PROTEIN FORMULATIONS AND METHODS OF MAKING SAME

• US 20090291062A1
• Filed: 11/28/2008
• Published: 11/26/2009
• Est. Priority Date: 11/30/2007
• Status: Active Grant

• Current Assignee: AbbVie Biotechnology Ltd. (Puerto Rico) (Abbvie Incorporated)
• Original Assignee: Abbvie Incorporated
• Inventors: FRAUNHOFER, Wolfgang, Krause, Hans-juergen, Kaleta, Katharina, Bartl, Annika, Tschoepe, Markus
• Granted Patent: US 8,420,081 B2
• Time in Patent Office: Days
• US Class Current: 424/85.5
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 38/46 Hydrolases (3)
• CPC Class Codes: A61K 38/4893 Botulinum neurotoxin (3.4.24.69)
• CPC Class Codes: A61K 38/50 acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
• 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: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39591 Stabilisation, fragmentation
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• 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 37/00 Drugs for immunological or allergic disorders

Abstract

The invention provides an aqueous formulation comprising water and a protein, and methods of making the same. The aqueous formulation of the invention may be a high protein formulation and/or may have low levels of conductivity resulting from the low levels of ionic excipients. Also included in the invention are formulations comprising water and proteins having low osmolality.

Claims

1. An aqueous formulation comprising a protein and water, wherein the formulation has a conductivity of less than about 2.5 mS/cm and the protein has a molecular weight (M_w) greater than about 47 kDa.

2. The formulation of claim 1, wherein the protein has a M_w greater than about 57 kDa.

3. The formulation of claim 1, wherein the protein has a M_w greater than about 100 kDa.

4. The formulation of claim 1, wherein the protein has a M_w greater than about 150 kDa.

6. The formulation of claim 1, wherein the protein has a M_w greater than about 250 kDa.

7. The formulation of claim 1, wherein the formulation has a conductivity of less than about 2 mS/cm.

9. The formulation of claim 1, wherein the formulation has a conductivity of less than about 1 mS/cm.

10. The formulation of claim 1, wherein the formulation has a conductivity of less than about 0.5 mS/cm.

31. The formulation of any one of claims 1, 11, 21, and 27, wherein the protein is an antibody, or an antigen-binding fragment thereof.

32. The formulation of claim 31, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

33. The formulation of claim 31, wherein the antibody, or antigen-binding fragment thereof, is an anti-TNFα

or an anti-IL-12 antibody.

34. The formulation of claim 31, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of Humira (adalimumab), Campath (Alemtuzumab), CEA-Scan Arcitumomab (fab fragment), Erbitux (Cetuximab), Herceptin (Trastuzumab), Myoscint (Imciromab Pentetate), ProstaScint (Capromab Pendetide), Remicade (Infliximab), ReoPro (Abciximab), Rituxan (Rituximab), Simulect (Basiliximab), Synagis (Palivizumab), Verluma (Nofetumomab), Xolair (Omalizumab), Zenapax (Daclizumab), Zevalin (Ibritumomab Tiuxetan), Orthoclone OKT3 (Muromonab-CD3), Panorex (Edrecolomab), Mylotarg (Gemtuzumab ozogamicin), golimumab (Centocor), Cimzia (Certolizumab pegol), Soliris (Eculizumab), CNTO 1275 (ustekinumab), Vectibix (panitumumab), Bexxar (tositumomab and 1131 tositumomab), and Avastin (bevacizumab).

35. The formulation of any one of claims 1, 11, 21, and 27, wherein the protein is a therapeutic protein.

36. The formulation of claim 35, wherein the therapeutic protein is selected from the group consisting of Pulmozyme (Dornase alfa), Rebif, Regranex (Becaplermin), Activase (Alteplase), Aldurazyme (Laronidase), Amevive (Alefacept), Aranesp (Darbepoetin alfa), Becaplermin Concentrate, Betaseron (Interferon beta-1b), BOTOX (Botulinum Toxin Type A), Elitek (Rasburicase), Elspar (Asparaginase), Epogen (Epoetin alfa), Enbrel (Etanercept), Fabrazyme (Agalsidase beta), Infergen (Interferon alfacon-1), Intron A (Interferon alfa-2a), Kineret (Anakinra), MYOBLOC (Botulinum Toxin Type B), Neulasta (Pegfilgrastim), Neumega (Oprelvekin), Neupogen (Filgrastim), Ontak (Denileukin diftitox), PEGASYS (Peginterferon alfa-2a), Proleukin (Aldesleukin), Pulmozyme (Dornase alfa), Rebif (Interferon beta-1a), Regranex (Becaplermin), Retavase (Reteplase), Roferon-A (Interferon alfa-2), TNKase (Tenecteplase), Xigris (Drotrecogin alfa), Arcalyst (Rilonacept), NPlate (Romiplostim), Mircera (methoxypolyethylene glycol-epoetin beta), Cinryze (C1 esterase inhibitor), Elaprase (idursulfase), Myozyme (alglucosidase alfa), Orencia (abatacept), Naglazyme (galsulfase), Kepivance (palifermin) and Actimmune (interferon gamma-1b).

42. The formulation of any one of claims 1, 11, 21, 27, and 37, further comprising a non-ionizable excipient.

43. The formulation of claim 42, wherein the non-ionizable excipient is selected from the group consisting of sugar alcohols and polyols such as mannitol or sorbitol, a non-ionic surfactant, sucrose, trehalose, raffinose, and maltose.

44. The formulation of claim 43, wherein the non-ionic surfactant is polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80.

45. The formulation of any one of claims 1, 11, 21, 27, and 37, wherein the formulation is stable in a liquid form for at least about 3 months or at least about 12 months.

48. The formulation of any one of claims 1, 11, 21, 27, and 37, wherein the formulation does not comprise an agent selected from the group consisting of a tonicity modifier, a stabilizing agent, a surfactant, an anti-oxidant, a cryoprotectant, a bulking agent, a lyroprotectant, a basic component, and an acidic component.

50. The formulation of any one of claims 1, 11, 21, 27, and 37, wherein the formulation is suitable for in vitro or in vivo use.

52. The formulation of claim 50, wherein the formulation is suitable for administration to a subject via a mode of administration selected from the group consisting of subcutaneous, intravenous, inhalation, intradermal, transdermal, intraperitoneal, and intramuscular.

53. Use of the formulation of claim 50 in the treatment of a disorder in a subject.

54. A device comprising the formulation of any one claims 1, 11, 21, 27, and 37.

56. An article of manufacture comprising the formulation of any one of claims 1, 11, 21, 27, and 37.

5. (canceled)

8. (canceled)

11. An aqueous formulation comprising a protein at a concentration of at least about 10 μ

g/mL and water, wherein the formulation has a conductivity of less than about 2 mS/cm.

13. The formulation of claim 11, wherein the concentration of the protein is at least about 10 mg/mL.

15. The formulation of claim 11, wherein the concentration of the protein is at least about 100 mg/mL.

16. The formulation of claim 11, wherein the concentration of the protein is at least about 150 mg/mL.

17. The formulation of claim 11, wherein the concentration of the protein is at least about 200 mg/mL.

18. The formulation of claim 11, wherein the concentration of the protein is greater than about 200 mg/mL.

20. The formulation of claim 11, wherein the formulation has a conductivity of less than about 0.5 mS/cm.

12. (canceled)

14. (canceled)

19. (canceled)

21. An aqueous formulation comprising a protein at a concentration of at least about 50 mg/mL and water wherein the formulation has an osmolality of no more than about 30 mOsmol/kg.

22. The formulation of claim 21, wherein the osmolality of the formulation is no more than about 15 mOsmol/kg.

24. The formulation of claim 21, wherein the concentration of the protein is at least about 150 mg/mL.

26. The formulation of claim 21, wherein the concentration of the protein is greater than about 200 mg/mL.

23. (canceled)

25. (canceled)

27. An aqueous formulation comprising water and a given concentration of a protein, wherein the protein has a hydrodynamic diameter (D_h) which is at least about 50% less than the D_h of the protein in a buffered solution at the given concentration.

28. The formulation of claim 27, wherein the D_h of the protein is at least about 50% less than the D_h of the protein in phosphate buffered saline (PBS) at the given concentration.

29. The formulation of claim 28, wherein the D_h of the protein is at least about 60% less than the D_h of the protein in PBS at the given concentration.

30. The formulation of claim 28, wherein the D_h of the protein is at least about 70% less than the D_h of the protein in PBS at the given concentration.

37. An aqueous formulation comprising an antibody, or an antigen-binding fragment, at a concentration of at least about 10 mg/mL and water, wherein the antibody, or antigen-binding fragment, has a hydrodynamic diameter (D_h) of less than about 5 μ

m.

38. The formulation of claim 37, wherein the antibody has a D_h of less than about 3 μ

m.

39. The formulation of claim 37, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

40. The formulation of claim 37, wherein the antibody, or antigen-binding fragment thereof, is an anti-TNFα

or an anti-IL-12 antibody.

41. The formulation of claim 37, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of Humira (adalimumab), Campath (Alemtuzumab), CEA-Scan Arcitumomab (fab fragment), Erbitux (Cetuximab), Herceptin (Trastuzumab), Myoscint (Imciromab Pentetate), ProstaScint (Capromab Pendetide), Remicade (Infliximab), ReoPro (Abciximab), Rituxan (Rituximab), Simulect (Basiliximab), Synagis (Palivizumab), Verluma (Nofetumomab), Xolair (Omalizumab), Zenapax (Daclizumab), Zevalin (Ibritumomab Tiuxetan), Orthoclone OKT3 (Muromonab-CD3), Panorex (Edrecolomab), and Mylotarg (Gemtuzumab ozogamicin), golimumab (Centocor), Cimzia (Certolizumab pegol), Soliris (Eculizumab), CNTO 1275 (ustekinumab), Vectibix (panitumumab), Bexxar (tositumomab and 1131 tositumomab) and Avastin (bevacizumab).

46. (canceled)

47. (canceled)

49. (canceled)

51. (canceled)

55. (canceled)

57. A method of preparing an aqueous formulation comprising a protein and water, the method comprising:

a) providing the protein in a first solution; and

b) subjecting the first solution to diafiltration using water as a diafiltration medium until at least a five fold volume exchange with the water has been achieved to thereby prepare the aqueous formulation.

60. The method of claim 57 or 58, wherein the diafiltration medium consists of water.

61. The method of claim 57 or 58, wherein the first solution is subjected to diafiltration with water until at least about a six-fold volume exchange or at least about a seven-fold volume exchange is achieved.

63. The method of claim 57 or 58, wherein the aqueous formulation has a final concentration of excipients which is at least about 95% less or 99% less than the first solution.

65. The method of claim 57 or 58, wherein the first protein solution is obtained from a mammalian cell expression system and has been purified to remove host cell proteins (HCPs).

66. The method of claim 57 or 58, wherein the protein has a M_w greater than about 1 kDa.

67. The method of claim 57 or 58, wherein the protein has a molecular weight (M_w) selected from the group consisting of greater than about 10 kDa, greater than about 47 kDa, greater than about 57 kDa, greater than about 100 kDa, greater than about 150 kDa, greater than about 200 kDa, and greater than about 250 kDa.

74. The method of claim 57 or 58, further comprising adding an excipient to the aqueous formulation.

75. The method of claim 74, wherein the aqueous formulation is a pharmaceutical formulation.

76. The method of claim 57 or 58, wherein the aqueous formulation is a pharmaceutical formulation.

77. The method of claim 76, further comprising loading the aqueous formulation into a device suitable for administering the aqueous formulation to a subject.

79. The method of claim 57 or 58, wherein the protein is an antibody, or an antigen-binding fragment thereof.

80. The method of claim 79, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of a chimeric antibody, a human antibody, a humanized antibody, and a domain antibody (dAb).

81. The method of claim 79, wherein the antibody, or antigen-binding fragment thereof, is an anti-TNF or an anti-IL-12 antibody.

82. The method of claim 79, wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of Humira (adalimumab), Campath (Alemtuzumab), CEA-Scan Arcitumomab (fab fragment), Erbitux (Cetuximab), Herceptin (Trastuzumab), Myoscint (Imciromab Pentetate), ProstaScint (Capromab Pendetide), Remicade (Infliximab), ReoPro (Abciximab), Rituxan (Rituximab), Simulect (Basiliximab), Synagis (Palivizumab), Verluma (Nofetumomab), Xolair (Omalizumab), Zenapax (Daclizumab), Zevalin (Ibritumomab Tiuxetan), Orthoclone OKT3 (Muromonab-CD3), Panorex (Edrecolomab), Mylotarg (Gemtuzumab ozogamicin), golimumab (Centocor), Cimzia (Certolizumab pegol), Soliris (Eculizumab), CNTO 1275 (ustekinumab), Vectibix (panitumumab), Bexxar (tositumomab and 1131 tositumomab), and Avastin (bevacizumab).

83. The method of claim 57 or 58, wherein the protein is a therapeutic protein.

84. The method of claim 83, wherein the therapeutic protein is selected from the group consisting of Pulmozyme (Dornase alfa), Rebif, Regranex (Becaplermin), Activase (Alteplase), Aldurazyme (Laronidase), Amevive (Alefacept), Aranesp (Darbepoetin alfa), Becaplermin Concentrate, Betaseron (Interferon beta-1b), BOTOX (Botulinum Toxin Type A), Elitek (Rasburicase), Elspar (Asparaginase), Epogen (Epoetin alfa), Enbrel (Etanercept), Fabrazyme (Agalsidase beta), Infergen (Interferon alfacon-1), Intron A (Interferon alfa-2a), Kineret (Anakinra), MYOBLOC (Botulinum Toxin Type B), Neulasta (Pegfilgrastim), Neumega (Oprelvekin), Neupogen (Filgrastim), Ontak (Denileukin diftitox), PEGASYS (Peginterferon alfa-2a), Proleukin (Aldesleukin), Pulmozyme (Dornase alfa), Rebif (Interferon beta-1a), Regranex (Becaplermin), Retavase (Reteplase), Roferon-A (Interferon alfa-2), TNKase (Tenecteplase), and Xigris (Drotrecogin alfa), Arcalyst (Rilonacept), NPlate (Romiplostim), Mircera (methoxypolyethylene glycol-epoetin beta), Cinryze (C1 esterase inhibitor), Elaprase (idursulfase), Myozyme (alglucosidase alfa), Orencia (abatacept), Naglazyme (galsulfase), Kepivance (palifermin) and Actimmune (interferon gamma-1b).

58. A method of preparing an aqueous formulation of a protein, the method comprising:

a) providing the protein in a first solution;

b) subjecting the first solution to diafiltration using water as a diafiltration medium until at least a five-fold volume exchange with the water has been achieved to thereby prepare a diafiltered protein solution; and

c) concentrating the diafiltered protein solution to thereby prepare the aqueous formulation of the protein.

59. The method of claim 58, wherein the concentration of the diafiltered protein solution is achieved via centrifugation.

86. An aqueous formulation prepared according to the methods of claim 58 or 59.

62. (canceled)

64. (canceled)

68-73. -73. (canceled)

78. (canceled)

85. (canceled)

87-101. -101. (canceled)