CRYSTALLINE ANTI-HUMAN IL-12 ANTIBODIES

• US 20120177704A1
• Filed: 02/14/2012
• Published: 07/12/2012
• Est. Priority Date: 03/29/2007
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Borhani, David W., Fraunhofer, Wolfgang, Krause, Hans-Juergen, Koenigsdorfer, Anette, Winter, Gerhard, Gottschalk, Stefan
• Granted Patent: US 8,404,819 B2
• Time in Patent Office: Days
• US Class Current: 424/400
• CPC Class Codes: A61K 39/39591 Stabilisation, fragmentation
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 15/10 for impotence
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 17/14 for baldness or alopecia
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/06 Antigout agents, e.g. antihyperuricemic or uricosuric agents
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 : for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• 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 3/00 : Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/08 : for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/14 : for RNA viruses
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 33/00 : Antiparasitic agents
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 5/16 : for decreasing, blocking or antagonising the activity of the thyroid hormones
• CPC Class Codes: A61P 5/38 : of the suprarenal hormones
• CPC Class Codes: A61P 5/40 : Mineralocorticosteroids, e.g. aldosterone; Drugs increasing or potentiating the activity of mineralocorticosteroids
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 16/244 : Interleukins [IL]
• CPC Class Codes: C07K 2299/00 : Coordinates from 3D structures of peptides, e.g. proteins or enzymes
• CPC Class Codes: C30B 7/00 : Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention relates to batch crystallization methods for crystallizing an anti-hIL-12 antibody that allows the production of the antibody on an industrial scale, antibody crystals obtained according to the methods, compositions containing the crystals, and methods of using the crystals and the compositions.

Claims

1. A batch crystallization method for crystallizing an anti-human IL-12 antibody, the method comprising the steps of:

(a) providing an aqueous solution of the antibody in admixture with at least one polyalkylene glycol as crystallization agent; and

(b) incubating the aqueous crystallization mixture until crystals of the antibody are formed.

2. The crystallization method according to claim 1, wherein the pH of the aqueous crystallization mixture is in the range of about pH 4 to about 6.5.

3. The crystallization method according to claim 1, wherein the aqueous crystallization mixture comprises a buffer.

4. The crystallization method according to claim 3, wherein the buffer comprises an acetate buffer.

5. The crystallization method according to claim 4, wherein the buffer comprises sodium acetate.

6. The crystallization method according to any one of claims 3 to 5, wherein the buffer concentration in the aqueous crystallization mixture is up to about 0.5 M.

7. The crystallization method according to any one of claims 1-5, wherein the polyalkylene glycol has an average molecular weight in the range of about 400 to about 10,000.

8. The crystallization method according to claim 7, wherein the polyalkylene glycol is polyethylene glycol.

9. The crystallization method according to any one of claims 1-5, wherein the polyalkylene glycol concentration in the crystallization mixture is in the range of about 5 to 30% (w/v).

10. The crystallization method according to claim 9, wherein the polyalkylene glycol is polyethylene glycol.

11. The crystallization method according to any one of claims 1-5, wherein at least one of the following additional crystallization conditions are met:

a) incubation is performed for between about 1 hour to about 250 days;

b) incubation is performed at a temperature between about 4°

C. and about 37°

C.;

c) the antibody concentration is in the range of about 0.5 to about 280 mg/ml.

12. The crystallization method according to claim 1, further comprising the step of drying the crystals.

13. The crystallization method according to any one of claims 1-5, further comprising the step of exchanging the crystallization mother liquor with an artificial mother liquor.

14. The crystallization method according to any one of claims 1-5, wherein the batch volume is in the range of about 1 ml to about 20,000 liters.

16. A crystal of an anti-human IL-12 antibody, obtainable by a crystallization method according to any one of claims 1-5.

41. The crystallization method according to any one of claims 1-5, further comprising the step of extending the yield of the crystals by adding additional polyalkylene glycol.

42. The method according to claim 41, wherein the polyalkylene glycol is polyethylene glycol.

43. The method according to claim 41, wherein the polyalkylene glycol is added continuously.

44. The crystallization method according to according to claim 1-5, further comprising the step of seeding the reaction with ABT-874.

15. A crystal of an anti-human IL-12 antibody.

17. The crystal according to claim 15, wherein the crystal has a sword-like morphology.

18. The crystal according to claim 15, wherein the antibody is a polyclonal antibody or a monoclonal antibody.

19. The crystal according to claim 18, wherein the antibody is selected from the group consisting of a chimeric antibody, a humanized antibody, a non-glycosylated antibody, a human antibody, and a mouse antibody.

20. The crystal according to claim 18, wherein the antibody is an IgG antibody.

21. The crystal according to claim 20, wherein the antibody is selected from the group consisting of an IgG1, an IgG2, an IgG3 and an IgG4 antibody.

22. The crystal according to claim 21, wherein the antibody is an anti-human IL-12 antibody of the group IgG1.

23. The crystal according to claim 22, wherein the crystal is prepared from an isolated human antibody that dissociates from human IL-12 with a Kd of 1×

10<sup>−

10</sup> M or less and a k_off rate constant of 1×

10<sup>−

3</sup> s<sup>−

1</sup> or less, both determined by surface plasmon resonance.

24. The crystal according to claim 22, wherein the crystal is prepared from an isolated human antibody with a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

2 and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO;

1.

25. The crystal according to claim 15, wherein the antibody is ABT-874.

26. A pharmaceutical composition comprising:

(a) crystals of an anti-human IL-12 antibody according to claim 15, and (b) at least one pharmaceutical excipient;

wherein the composition is provided as a solid, a semisolid or a liquid formulation, each formulation containing the antibody in crystalline form.

27. A pharmaceutical composition comprising:

(a) crystals of an anti-human IL-12 antibody according to claim 15, and (b) at least one pharmaceutical excipient, which embeds or encapsulates the crystals.

28. The composition of claim 26 or 27, wherein the composition has an antibody concentration greater than about 1 mg/ml.

29. The composition of claim 28, wherein the composition has an antibody concentration greater than about 200 mg/ml.

30. The composition according to claims 26 and 27, wherein the composition comprises at least one carrier selected from the group consisting of a polymeric biodegradable carrier, a polymeric non-biodegradable carrier, an oil carrier, and a lipid carrier.

31. The composition according to claim 30, wherein the polymeric carrier is a polymer selected from one or more of the group consisting of:

poly(acrylic acid), poly(cyanoacrylates), poly(amino acids), poly(anhydrides), poly(depsipeptide), poly (esters), poly(lactic acid), poly(lactic-co-glycolic acid) or PLGA, poly(β

-hydroxybutryate), poly(caprolactone), poly(dioxanone);

poly(ethylene glycol), poly(hydroxypropyl)methacrylamide, poly[(organo)phosphazene, poly(ortho esters), poly(vinyl alcohol), poly(vinylpyrrolidone), maleic anhydride alkyl vinyl ether copolymers, pluronic polyols, albumin, alginate, cellulose and cellulose derivatives, collagen, fibrin, gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans, sulfated polysaccharides, blends and copolymers thereof.

32. An injectable liquid composition comprising anti-human IL-12 antibody crystals according to claim 15 and having an antibody concentration in the range of about 10 to about 400 mg/ml.

33. A crystal slurry composition comprising anti-human IL12 antibody crystals according to claim 15, having an antibody concentration greater than about 100 mg/ml.

34. A method for treating a mammal comprising the step of administering to the mammal an effective amount of anti-human IL-12 antibody crystals according to claim 15.

35. A method for treating a mammal comprising the step of administering to the mammal an effective amount of the composition according to claim 26 or 27.

36. The method according to claim 35, wherein the composition is administered by a parenteral route, an oral route, or by an injection.

37. A method of treating an IL-12-related disorder in a subject, which method comprises administering a therapeutically effective amount of the antibody crystals according to claim 15.

38. The method of claim 37, wherein the IL-12-related disorder is selected from the group consisting of rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn'"'"'s disease, ulcerative colitis, inflammatory bowel disease, insulin dependent diabetes mellitus, thyroiditis, asthma, allergic diseases, psoriasis, dermatitis scleroderma, atopic dermatitis, graft versus host disease, organ transplant rejection, acute or chronic immune disease associated with organ transplantation, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki'"'"'s disease, Grave'"'"'s disease, nephrotic syndrome, chronic fatigue syndrome, Wegener'"'"'s granulomatosis, Henoch-Schoenlein purpurea, microscopic vasculitis of the kidneys, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome, sepsis syndrome, cachexia, infectious diseases, parasitic diseases, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington'"'"'s chorea, Parkinson'"'"'s disease, Alzheimer'"'"'s disease, stroke, primary biliary cirrhosis, hemolytic anemia, malignancies, heart failure, myocardial infarction, Addison'"'"'s disease, sporadic, polyglandular deficiency type I and polyglandular deficiency type II, Schmidt'"'"'s syndrome, adult (acute) respiratory distress syndrome, alopecia, alopecia greata, seronegative arthopathy, arthropathy, Reiter'"'"'s disease, psoriatic arthropathy, ulcerative colitic arthropathy, enteropathic synovitis, chlamydia, yersinia and salmonella associated arthropathy, spondyloarthopathy, atheromatous disease/arteriosclerosis, atopic allergy, autoimmune bullous disease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid, linear IgA disease, autoimmune haemolytic anaemia, Coombs positive haemolytic anaemia, acquired pernicious anaemia, juvenile pernicious anaemia, myalgic encephalitis/Royal Free Disease, chronic mucocutaneous candidiasis, giant cell arteritis, primary sclerosing hepatitis, cryptogenic autoimmune hepatitis, Acquired Immunodeficiency Disease Syndrome, Acquired Immunodeficiency Related Diseases, Hepatitis C, common varied immunodeficiency (common variable hypogammaglobulinaemia), dilated cardiomyopathy, female infertility, ovarian failure, premature ovarian failure, fibrotic lung disease, cryptogenic fibrosing alveolitis, post-inflammatory interstitial lung disease, interstitial pneumonitis, connective tissue disease associated interstitial lung disease, mixed connective tissue disease associated lung disease, systemic sclerosis associated interstitial lung disease, rheumatoid arthritis associated interstitial lung disease, systemic lupus erythematosus associated lung disease, dermatomyositis/polymyositis associated lung disease, Sjodgren'"'"'s disease associated lung disease, ankylosing spondylitis associated lung disease, vasculitic diffuse lung disease, haemosiderosis associated lung disease, drug-induced interstitial lung disease, radiation fibrosis, bronchiolitis obliterans, chronic eosinophilic pneumonia, lymphocytic infiltrative lung disease, postinfectious interstitial lung disease, gouty arthritis, autoimmune hepatitis, type-1 autoimmune hepatitis (classical autoimmune or lupoid hepatitis), type-2 autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune mediated hypoglycemia, type B insulin resistance with acanthosis nigricans, hypoparathyroidism, acute immune disease associated with organ transplantation, chronic immune disease associated with organ transplantation, osteoarthrosis, primary sclerosing cholangitis, idiopathic leucopenia, autoimmune neutropenia, renal disease NOS, glomerulonephritides, microscopic vasulitis of the kidneys, lyme disease, discoid lupus erythematosus, male infertility idiopathic or NOS, sperm autoimmunity, multiple sclerosis (all subtypes), insulin-dependent diabetes mellitus, sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, Goodpasture'"'"'s syndrome, pulmonary manifestation of polyarteritis nodosa, acute rheumatic fever, rheumatoid spondylitis, Still'"'"'s disease, systemic sclerosis, Takayasu'"'"'s disease/arteritis, autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune thyroid disease, hyperthyroidism, goitrous autoimmune hypothyroidism (Hashimoto'"'"'s disease), atrophic autoimmune hypothyroidism, primary myxoedema, phacogenic uveitis, primary vasculitis and vitiligo. The human antibodies, and antibody portions of the invention can be used to treat autoimmune diseases, in particular those associated with inflammation, including, rheumatoid spondylitis, allergy, autoimmune diabetes, autoimmune uveitis.

39. The use of anti-human IL-12 antibody crystals according to claim 15 for preparing a pharmaceutical composition for treating an IL-12-related disease.

40. Anti-human IL-12 antibody crystals according to claim 15 for use in medicine.