HIGH AFFINITY AND AGGREGATIVELY STABLE ANTIBODIES ON THE BASIS OF VARIABLE DOMAINS VL AND A DERIVATIVE VHH

• US 20170066843A1
• Filed: 09/01/2016
• Published: 03/09/2017
• Est. Priority Date: 09/26/2014
• Status: Active Grant

• Current Assignee: Closed Joint-Stock Company "BIOCAD
• Original Assignee: Closed Joint-Stock Company "BIOCAD
• Inventors: ULITIN, Andrey Borisovich, EVDOKIMOV, Stanislav Rudolfovich, SOLOVIEV, Valeriy Vladimirovich, CHERNYH, Yulia Sergeevna, GONCHAROVA, Olga Vladimirovna, KORZHAVIN, Dmitriy Valerievich, CHERNOVSKAYA, Tatyana Veniaminovna, NEMANKIN, Timofey Aleksandrovich, IVANOV, Roman Alexeevich, MOROZOV, Dmitriy Valentinovich, EKIMOVA, Victoria Mikhailovna, SOFRONOVA, Ekaterina Vladimirovna, USTYUGOV, Yakov Yurevich
• Granted Patent: US 10,442,869 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/08 of the prostate
• 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 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/04 Antipruritics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/04 : Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 25/06 : Antimigraine agents
• CPC Class Codes: A61P 25/10 : for petit-mal
• 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 25/18 : Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/22 : Anxiolytics
• CPC Class Codes: A61P 25/24 : Antidepressants
• 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 25/30 : for treating abuse or dependence
• CPC Class Codes: A61P 25/32 : Alcohol-abuse
• 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 3/08 : for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/06 : for tuberculosis
• CPC Class Codes: A61P 31/10 : Antimycotics
• CPC Class Codes: A61P 31/14 : for RNA viruses
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 31/20 : for DNA viruses
• CPC Class Codes: A61P 33/02 : Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• 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 5/18 : of the parathyroid hormones
• 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 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• 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 9/06 : Antiarrhythmics
• CPC Class Codes: A61P 9/08 : Vasodilators for multiple indications
• 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: C07K 16/244 : Interleukins [IL]
• CPC Class Codes: C07K 16/461 : Igs containing Ig-regions, -domains or -residues form different species
• CPC Class Codes: C07K 2317/22 : from camelids, e.g. camel, llama or dromedary
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/567 : Framework region [FR]
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

The monoclonal IgG-type antibodies were suggested comprising variable domains represented by a combination of VHH-derivative with a variable domain of the light chain V_L. Said antibodies can comprise amino acid substitutions at positions 44 and 45 (Kabat numbering) or combinations thereof. Antibodies of the invention possess increased affinity and improved aggregation stability.

Claims

1. A humanized monoclonal IgG antibody or a fragment thereof wherein the variable domains are represented by a combination of a V_HH-derivative with a variable domain of the light chain V_L.

2. The antibody or the fragment thereof according to claim 1, wherein the V_HH-derivative comprises the following amino acid substitutions at positions 44X₂45 X₃:

a) wherein 44X2=G, A, V, S, T;

b) wherein 45X3=A, V, T, H;

or combinations thereof.

3. The antibody or the fragment thereof according to claim 1, characterized by increased aggregation stability of V_HH-derivative compared to the native VHH antibody isolated from an immunized animal from Camelidae family.

4. The antibody or the fragment thereof according to claim 1, wherein the V_HH-derivative is represented by:

a variable domain of the heavy chain isolated from an immunized animal from Camelidae family;

or

a variable domain of the heavy chain isolated from a non-immunized animal from Camelidae family;

and comprises additional amino acid substitutions typical for humans and located at any positions, except for the following substitutions at positions 44X₂45 X₃;

c) wherein 44X2=G, A, V, S, T;

d) wherein 45X3=A, V, T, H;

or combinations thereof.

5. The antibody or the fragment thereof according to claim 1, wherein the variable domain of the light chain V_L is a human antibody derivative or a humanized fragment of a mammalian antibody.

6. The antibody according to claim 1, wherein said antibody is IgG1, IgG2, IgG3 or IgG4.

7. The antibody according to claim 6, comprising a non-native modified Fc as a part of IgG.

8. The antibody or the fragment thereof according to claim 1, wherein said antibody has at least one of the following properties:

a) an aggregation stability such that when used in concentrations over 10 mg/ml and stored for >

6 months at a temperature of 4°

C., the content of aggregates increases by not more than 5% of their initial content in the solution;

b) an aggregation stability such that when used in concentrations over 10 mg/ml and stored for >

2 weeks at a temperature of 37°

C., the content of aggregates increases by not more than 5% of their initial content in the solution;

c) an aggregation stability such that when used in concentrations over 10 mg/ml and stored for >

48h at a temperature of 42°

C., the content of aggregates increases by not more than 5% of their initial content in the solution;

d) an aggregation stability such that when used in concentrations over 10 mg/ml and stored for >

6 h at a temperature of 50°

C., the content of aggregates increases by not more than 5% of their initial content in the solution;

e) a dissociation constant K_D of 10<sup>−

9</sup> M or lower;

f) an antibody-antigen interaction kinetic association constant k_on(l/Ms) of at least 10⁵ l/Msec or more;

or

g) an antibody-antigen interaction kinetic dissociation constant dis(Vs) of 10⁴ l/sec or lower.

9. The antibody or the fragment thereof according to claim 1 that specifically binds to human IL-17A, comprising

a) a derivative of Camelidae heavy chain variable domain (V_HH) that comprises 3 hypervariable regions HCDR1, HCDR2 and HCDR3, wherein:

HCDR1 comprises the amino acid sequence of SEQ ID NO;

1;

G-T-F-A-T-X32-X33-X34-X35, wherein

X32 is an amino acid selected from the group consisting of S, N, K, R, E, W, Q, D, A, V and F;

X33 is an amino acid selected from the group consisting of P and S;

X34 is an amino acid selected from the group consisting of M and I; and

X35 is an amino acid selected from the group consisting of G, N, S, A, L, I, R, V and Q;

HCDR2 comprises the amino acid sequence of SEQ ID NO;

2;

X50-I-X52-X52a-S-G-X55-D-R-I-Y-A-D-S-V-K-G, wherein

X50 is an amino acid selected from the group consisting of A, G and L;

X52 is an amino acid selected from the group consisting of S, D and E;

X52a is an amino acid selected from the group consisting of P and A; and

X55 is an amino acid selected from the group consisting of G, S, T, L, R, D, E, K, A and W;

HCDR3 comprises the amino acid sequence of SEQ ID NO;

3;

C-A-X94-X95-X96-X97-F-X99-X100-X100a-X100b-X100c-X100d-X100e-X100f-D-Y-D-S, wherein

X94 is an amino acid selected from the group consisting of K, S, T, V, D and G;

X95 is an amino acid selected from the group consisting of R and K;

X96 is an amino acid selected from the group consisting of G, R, Y, H, D, W and K;

X97 is an amino acid selected from the group consisting of R, A, V, S, L and H;

X99 is an amino acid selected from the group consisting of D, E, G, A, R, V, K and Q;

X100 is an amino acid selected from the group consisting of G, S and N;

X100a is an amino acid selected from the group consisting of G, T, P, V, R, N and K;

X100b is an amino acid selected from the group consisting of V, S, T, L, Y, A, H, G and I;

X100c is an amino acid selected from the group consisting of Y, W and S;

X100d is an amino acid selected from the group consisting of R, V, L, Y, A, W, K, G, Q and I;

X100e is an amino acid selected from the group consisting of T, L, A and S; and

X100f is an amino acid selected from the group consisting of T, L, G, P, N, A, Q, F, I and D; and

b) a variable domain of the light chain (V_L) of a human antibody or a variable domain of the light chain of a humanized antibody.

10. The antibody or the fragment thereof according to claim 1 that specifically binds to human IL-17A, comprising:

a) a derivative of the heavy chain variable domain (V_HH) comprising 3 hypervariable regions HCDR1, HCDR2 and HCDR3, wherein;

HCDR1 comprises the amino acid sequence of G-T-F-A-T-S-P-M-G (SEQ ID NO;

     4);

HCDR2 comprises the amino acid sequence of A-I-S-P-S-G-G-D-R-I-Y-A-D-S-V-K-G (SEQ ID NO;

     5); and

HCDR3 comprises the amino acid sequence of C-A-V-R-R-R-F-D-G-T-S-Y-Y-T-G-D-Y-D-S(SEQ ID NO;

     6); and

b) a variable domain of the light chain (V_L) of a human antibody or a variable domain of the light chain of a humanized antibody.

11. The antibody or the fragment thereof according to claim 10 that specifically binds to human IL-17A, comprising:

a) a derivative of the heavy chain variable domain (V_HH) comprising the amino acid sequence

QVQLVQSGGGLVQAGGSLRLSCAASGGTFATSPMGWLRQAPGKGTEFVAAI SPSGGDRIYADSVKGRFTISRDNAGYFIYLQMNSLKPEDTAVYYCAVRRRFDGTSYY TGDYDSWGQGTLVTVSS (SEQ ID NO;

     7); and

b) a variable domain of the light chain (V_L) of a human antibody or a variable domain of the light chain of a humanized antibody.

12. The antibody or the fragment thereof according to claim 11 that specifically binds to human IL-17A, wherein said derivative of said V_HH comprises an amino acid sequence at least 90% identical to SEQ ID NO:

7.

13. The antibody or the fragment thereof according to claim 12 that specifically binds to human IL-17A, wherein said variable domain of said light chain (V_L) comprises the amino acid sequence

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYDASS RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYSYSPVTFGQGTKVEIKR (SEQ ID NO:

8).

14. The antibody or the fragment thereof according to claim 13 that specifically binds human IL-17A, wherein said variable domain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

8.

15. The antibody or the fragment thereof according to claim 1 wherein the antibody or the fragment thereof specifically binds human IL-17A and wherein:

a) the heavy chain comprises the following amino acid sequence;

QVQLVQSGGGLVQAGGSLRLSCAASGGTFATSPMGWLRQAPGKGTEFVAAI SPSGGDRIYADSVKGRFTISRDNAGYFIYLQMNSLKPEDTAVYYCAVRRRFDGTSYY TGDYDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKR VEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK (SEQ ID NO;

     9); and

b) the variable domain of the light chain (V_L) comprises the following amino acid sequence;

16. The antibody or the fragment thereof according to claim 15 that specifically binds human IL-17A, wherein said heavy chain comprises an amino acid sequence at least 90% identical to SEQ ID NO:

9 and/or said variable domain of said V_L comprises an amino acid sequence at least 90% identical to SEQ ID NO;

10.

17. An antibody or the fragment thereof according to claim 9, wherein said antibody has at least one of the following properties:

a) a binding affinity to human IL-17A characterized by K_D of 10<sup>−

10</sup> M or lower;

b) a kinetic association constant k_on(l/Mc) for human IL-17A of not less than 10⁵ l/Msec;

or

c) a kinetic dissociation constant dis(Vs) for human IL-17A of not more than 10<sup>−

5</sup> l/sec.

d) inhibits the activity of human IL-17A by no less than 50%.

18. The antibody or the fragment thereof according to claim 9 comprising one or more additional amino acid substitutions in the Fc-region compared to the native Fc, wherein said substitutions improve physical-chemical and pharmacokinetic properties of the antibody, as compared to an antibody with the native Fc, and do not reduce the antibody'"'"'s ability to bind IL-17A.

19. A DNA construct encoding the antibody or the fragment thereof according to claim 1.

20. An expression vector comprising the DNA construct according to claim 19.

21. A host cell comprising the expression vector according to claim 20.

22. A method for producing the antibody or the fragment thereof according to claim 1, which involves incubation of a host cell according to claim 21 in a culture medium under the conditions sufficient to obtain said antibody or fragment thereof, followed by isolation and purification of obtained antibody or its active fragment.

23. A pharmaceutical composition comprising the antibody or the fragment thereof according to claim 1, and one or more pharmaceutically suitable excipient, diluent or carrier.

24. The pharmaceutical composition according to claim 23, further comprising an active pharmaceutical ingredient which is a TNF-α

inhibitor.

25. A method for treatment of an IL-17A-mediated disease or disorder, comprising administration of a therapeutically effective amount of the antibody or the fragment thereof according to claim 1 to a patient in need thereof.

26. The method according to claim 25, wherein the IL-17A-mediated disease or disorder is selected from:

rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn'"'"'s disease, ulcerative colitis, inflammatory bowel disease, insulin dependent diabetes mellitus, thyroadenitis, asthma, allergic disorders, psoriasis, dermatitis, systemic sclerosis, graft-versus-host disease, graft rejection, acute or chronic immune disease associated with organ grafting, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki disease, Graves'"'"' disease, nephrotic syndrome, chronic fatigue syndrome, Wegener'"'"'s disease, Henoch-Schonlein purpura, microscopic polyangiitis with renal involvement, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome, sepsis syndrome, cachexia, infections, invasions, acquired immune deficiency syndrome, acute transverse myelitis, Huntington chorea, Parkinson disease, Alzheimer disease, stroke, primary biliary cirrhosis, hemolytic anemia, malignancies, heart failure, myocardial infarction, Addison disease, polyglandular autoimmune syndrome type I and type II, Schmidt'"'"'s syndrome, acute respiratory distress syndrome, alopecia, alopecia areata, seronegative arthropathy, arthropathy, Reiter'"'"'s syndrome, psoriatic arthropathy, arthropathy associated with ulcerative colitis, enteropathic synovitis, arthropathy associated with Chlamydia, Yersinia and Salmonella, spondyloarthropathy, atheromatosis disease/coronary sclerosis, atopic allergy, autoimmune bullous disease, pemphigus, pemphigus foliaceus, pemphigoid, linear IgA diseases, autoimmune hemolytic anemia, Coombs positive hemolytic anemia, acquired pernicious anemia, juvenile pernicious anemia, Myalgic encephalomyelitis/chronic fatigue syndrome, chronic active hepatic inflammation, cranial giant arteritis, primary sclerosing hepatitis, cryptogenic autoimmune hepatitis, acquired immune deficiency syndrome (AIDS), AIDS-associated diseases, hepatitis B, hepatitis C, common variable immunodeficiency (common variable hypogammaglobulinemia), dilated cardiomyopathy, female sterility, ovarian insufficiency, Premature ovarian failure, pulmonary fibrosis, cryptogenic fibrosing alveolitis, post inflammatory interstitial lung pathologies, interstitial pneumonitis, connective tissue disease associated with interstitial lung disease, mixed connective tissue disease associated with interstitial lung disease, systemic scleroderma associated with interstitial lung disease, rheumatoid arthrisit associated with interstitial lung disease, systemic lupus erythematosus associated with lung disease, dermatomyositis/polymyositis associated with lung disease, Sjogren disease associated with lung disease, ankylosing spondylitis associated with lung disease, diffuse pulmonary vasculitis, hemosiderosis associated with lung disease, drug-induced interstitial lung disease, fibrosis, radiation-induced fibrosis, obliterating bronchiolitis, chronic eosinophilic pneumonia, lung disease with lymphocyte infiltration, post infectious interstitial lung pathologies, gouty arthritis, autoimmune hepatitis, autoimmune hepatitis type I (classic autoimmune or lupoid hepatitis), autoimmune hepatitis type II (associated with anti-LKM antibody), autoimmune hypoglycemia, type B insulin resistance with acanthokeratodermia, hypoparathyroidism, acute graft-associated immune disease, chronic graft-associated immune disease, osteoarthrosis, primary sclerosing cholangitis, type I psoriasis, type II psoriasis, idiopathic leukopenia, autoimmune neutropenia, NOS-kidney diseases, glomerulonephritis, microscopic renal polyangiitis, Lyme disease, discoid lupus erythematosus, idiopathic of NOS-male sterility, antisperm immunity, multiple sclerosis (all types), sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, Goodpasture syndrome, pulmonary manifestations of polyarteritis nodosa, acute rheumatic fever, rheumatoid spondylitis, Still'"'"'s disease, systemic scleroderma, Sjogren'"'"'s Syndrome, Takayasu disease/arthritis, autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune thyroid disorders, hyperthyroid, autoimmune hypothyroidism (Hashimoto disease), atrophic autoimmune hypothyroidism, primary myxedema, phacogenic uveitis, primary vasculitis, vitiligo, acute hepatic disease, chronic hepatic disease, alcoholic cirrhosis, alcohol-induced liver damage, cholestasis, idiosyncratic hepatic disease, drug-induced hepatitis, nonalcoholic steatohepatitis, allergies and asthma, group B streptococcal infection (GBS), mental disorders (including depressions and schizophrenia), Th1- and Th2-mediated disease, acute and chronic pain (various forms), malignancies such as lung cancer, breast cancer, stomach cancer, bladder cancer, colorectal cancer, pancreatic cancer, ovarian cancer, prostate cancer and hematopoietic malignancies (leukemia and lymphomas), abetalipoproteinaemia, acrocyanosis, acute and chronic infections and infestations, acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute or chronic bacterial infection, acute pancreatitis, acute renal failure, adenocarcinoma, atrial ectopics, AIDS dementia complex, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, allograft rejection, alpha-I antitrypsin deficiency, lateral amyotrophic sclerosis, anemia, angina, anterior horn cell degeneration, anti-CD3 therapy, antiphospholipid syndrome, hypersensitivity reactions against receptors, aortic and peripheral aneurysms, aortic dissection, arterial hypertension, coronary sclerosis, arteriovenous fistula, ataxia, atrial fibrillation (constant or paroxysmal), atrial flutter, atrioventricular block, B-cell lymphoma, bone graft rejection, bone marrow transplant (BMT) rejection, bundle branch block, Burkitt lymphoma, burns, cardiac arrythmia, myocardial stunning syndrome, cardiac tumor, cardiomyopathy, inflammatory response to bypass, cartilage graft rejection, brain cortex degeneration, cerebellar disorder, chaotic or multifocal atrial tachycardia, chemotherapy-induced disorders, chronic myelocytic leukemia (CML), chronic alcohol addiction, chronic inflammatory pathologies, chronic lymphatic leukemia (CLL), chronic obstructive pulmonary disease, chronic salicylate intoxication, rectocolic carcinoma, congestive cardiac failure, conjunctivitis, contact dermatitis, pulmonary heart, coronary artery disease, Creutzfeldt-Jakob Disease, culture-negative sepsis, cystic fibrosis, cytokine therapy-induced disorders, boxer'"'"'s encephalopathy, demyelinating disease, dengue hemorrhagic fever, dermatitis, dermatological conditions, diabetes, diabetes mellitus, diabetes-related atherosclerotic vascular disease, diffuse Lewy body disease, congestive dilated cardiomyopathy, basal ganglia disease, Down'"'"'s syndrome in middle age, motor disorders induced by CNS dopamine blockers, drug sensitivity, eczema, encephalomyelitis, endocarditis, endocrinopathy, epiglottiditis, Epstein-Barr viral infection, erythralgia, extrapyramidal and cerebellar symptoms, familial hemophagocytic lymphohistiocytosis, fetal thymus graft rejection, Friedreich'"'"'s ataxia, peripheral artery disease, fungal sepsis, gas phlegmon, gastric ulcer, glomerulonephritis, any organ or tissue graft rejection, gram negative sepsis, gram positive sepsis, granulomas due to intracellular organisms, hairy-cell leukemia, Hallervorden-Spatz disease, Hashimoto'"'"'s thyroiditis, hay fever, heart transplant rejection, hemochromatosis, hemodialysis, hemolytic uremic syndrome/thrombotic thrombocytopenic purpura, bleeding, hepatitis (A), bundle branch arrhythmia, HIV-infections/HIV-neuropathies, Hodgkin disease, hyperkinetic motor disorders, hypersensitivity reactions, hypersensitivity-associated pneumonitis, hypertension, hypokinetic motor disorders, examination of hypothalamo-pituitary-adrenal axis, idiopathic Addison'"'"'s disease, idiopathic pulmonary fibrosis, antibody-mediated cytotoxicity, asthenia, infantile muscular atrophy, aortal inflammation, influenza virus A, exposure to ionizing radiation, iridocyclitis/uveitis/optic neuritis, ischaemia/reperfusion-induced disorders, ischaemic stroke, juvenile rheumatoid arthritis, juvenile spinal muscular atrophy, Kaposi'"'"'s sarcoma, renal transplant rejection, legionellosis, leishmaniasis, leprosy, corticospinal damage, lipoedema, liver transplant rejection, lymphoedema, malaria, malignant lymphoma, malignant histiocytosis, malignant melanoma, meningitis, meningococcemia, metabolic/idiopathic diseases, migraine, multiple system mitochondrial disorders, mixed connective-tissue disease, monoclonal gammapathy, multiple myeloma, multiple system degeneration (Mencel Dejerine-Thomas Shi-Drager and Machado-Joseph), myasthenia gravis, intracellular Mycobacterium avium, Mycobacterium tuberculosis, myelodysplastic syndrome, myocardial infarction, myocardial ischemic disease, nasopharyngeal cancer, neonatal chronic lung disease, nephritis, nephrotic, neurodegenerative disorders, neurogenic muscular atrophy I, neutropenic fever, non-Hodgkin'"'"'s lymphomas, abdominal aortic branch occlusion, arterial occlusive disease, OKT3®

treatment, orchitis/epididymitis, orchitis/vasectomy reversal operations, organomegaly, osteoporosis, pancreatic graft rejection, pancreatic carcinoma, paraneoplastic disease/tumor-related hypercalcemia, parathyroid graft rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherosclerosis (atherlosclerotic) disease, peripheral vascular disease, peritonitis, pernicious anemia, Pneumocystis carinii pneumonia, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal plasma-proliferative disorder and skin changes), postperfusion syndrome, pump head syndrome, post-cardiotomy post-infarction syndrome, preeclampsia, progressive supranuclear paralysis, primary pulmonary hypertension, radiation therapy, Raynaud'"'"'s phenomenon and disease, Raynoud'"'"'s disease, Refsum'"'"'s disease, regular narrow QRS tachycardia, renal vascular hypertension, reperfusion injury, restrictive cardiomyopathy, sarcoma, scleroderma disease, senile chorea, Dementia with Lewy bodies, seronegative arthritis, shock, sickle cell disease, skin allograft rejection, skin changes, small intestinal graft rejection, solid tumors, specific arrhythmias, spinal ataxia, spinocerebellar degradations, streptococcal myositis, cerebellar structural damage, subacute sclerosing panencephalitis, syncope, cardiovascular syphilis, systemic anaphylaxis, a comprehensive systemic inflammatory response syndrome, systemic-onset juvenile rheumatoid arthritis, T cells or FAB ALL, telangiectasia, thrombosis obliterans, thrombocytopenia, toxicity, grafting, trauma/bleeding, hypersensitivity reactions type III, hypersensitivity reactions type IV, unstable angina, uremia, urinary sepsis, urticaria, valvular heart disease, varicose veins, vasculitis, venous diseases, venous thrombosis, ventricular fibrillation, viral and fungal infections, vital encephalitis/aseptic meningitis, vital hemophagocytic syndrome, Wernicke-Korsakoff syndrome, Wilson'"'"'s disease, heterograft rejection for any organ or tissue, acute coronary syndrome, acute idiopathic polyneuritis, acute inflammatory demyelinating radicular neuropathy, acute ischemia, adult-onset Stills disease, alopecia areata, anaphylaxis, antiphospholipid antibody syndrome, aplastic anemia, coronary sclerosis, atopic eczema, atopic dermatitis, autoimmune dermatitis, autoimmune disorder associated with streptococcus infection, autoimmune enteropathy, autoimmune hearing loss, autoimmune lymphoproliferative syndrome (ALPS), autoimmune myocarditis, autoimmune premature ovarian failure, blepharitis, bronchiectasis, bullous pemphigoid, cardiovascular disease, catastrophic antiphospholipid syndrome, celiac disease, cervical spondylosis, chronic ischemia, cicatrical pemphigoid, clinically isolated syndrome (cis) with the risk for multiple sclerosis, conjunctivitis, childhood-onset mental disorders, chronic obstructive pulmonary disease (COPD), dacryocystitis, dermatomyositis, diabetic retinopathy, diabetes mellitus, herniated disk, prolapse of intervertebral disc, drug-induced immune haemolytic anaemia, endocarditis, endometreosis, entophthalmia, episcleritis, erythema multiform, severe erythema multiform, gestational pemphigoid, Guillain-Barre syndrome (GBS), hay fever, Hughes syndrome, idiopathic Parkinson'"'"'s disease, idiopathic interstitial pneumonia, IgE-mediated allergy, autoimmune hemolytic anemia, inclusion body myositis, infectious ocular inflammatory disease, inflammatory demyelinating disease, inflammatory heart disease, inflammatory kidney disease, idiopathic pulmonary fibrosis/usual interstitial pneumonia, iritis, keratitis, keratoconjunctivitis sicca, Kussmaul disease or Kussmaul-Meier Disease, Landry palsy, Langerhans'"'"' cell histiocytosis, marbled skin, macular degeneration, microscopic polyangiitis, Bechterew disease, motor neuron disease, mucosal pemphigoid, multiple organ failure, myasthenia gravis, spinal cord dysplasia syndrome, myocarditis, nerve root disorders, neuropathy, non-A, non-B hepatitis, optic neuritis, osteolysis, ovarian cancer, oligoarticular JIA, peripheral arterial occlusive disease, peripheral vascular disease, peripheral artery disease (PAD), phlebitis, polyarteritis nodosa, polychondritis, polymyalgia rheumatica, poliosis, polyarticular juvenile idiopathic arthritis, multiple endocrine deficience, polymyositis, polymyalgia rheumatica (PMR), post pump syndrome, primary parkinsonism, prostate and rectal cancer and hematopoietic malignancies (leukemia and lymphoma), prostatitis, pure red-cell aplasia, primary adrenal insufficiency, relapsing neuromyelitis optica, restenosis, rheumatic heart disease, SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis), scleroderma, secondary amyloidosis, shock lung, scleritis, ischias, secondary adrenal insufficiency, silicon-associated connective tissue disease, Sneddon-Wilkinson disease, ankylosing spondylitis, Stevens-Johnson syndrome, systemic inflammation response syndrome, cranial arteritis, Toxoplasma rhinitis, toxic epidermal necrolysis, transverse myelitis, TRAPS (tumor necrosis factor receptor-associated periodic syndrome), allergic reactions type I, diabetes type II, urticaria, usual interstitial pneumonia, vasculitis, vernal conjunctivitis, viral retinitis, Vogt-Koyanagi-Harada syndrome (VKH syndrome), wet macular degeneration, wound healing, and Yersinia- or Salmonella-associated arthropathy.

27. The method according to 26, further comprising the administration of a TNF-α

inhibitor.