Anti-TNF antibodies, compositions, and methods for treatment of active ankylosing spondylitis

Abstract

To provide a method for treating active ankylosing spondylitis in a patient.SOLUTION: The present invention provides a method that includes administering to a patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising a specific amino acid sequence and a light chain (LC) comprising another specific amino acid sequence.SELECTED DRAWING: None

Description

Technical Field

[0001] (Reference to Electronically Submitted Sequence Listing) This application includes an ASCII-formatted sequence listing entitled "JBI6104WOPCT1Seq Listing.txt" created on May 4, 2020, having a size of 21 KB, which was electronically submitted via EFS-W eb. The sequence listing submitted via EFS-W eb is part of this specification and is hereby incorporated by reference in its entirety into this specification.

[0002] (Field of the Invention) The present invention relates to the treatment of ankylosing spondylitis (AS), for example, in the treatment using an anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, and to compositions and methods using an anti-TNF antibody or an antigen-binding fragment thereof.

Background Art

[0003] TNFα is a soluble homotrimer of a 17 kD protein subunit. A membrane-bound 26 kD precursor form of TNF also exists.

[0004] Cells other than monocytes or macrophages also produce TNFα. For example, human non-monocytic tumor cell lines produce TNFα as well as CD4+ and CD8+ peripheral blood T lymphocytes, and some cultured T and B cell lines also produce TNFα.

[0005] TNFα induces degradation of cartilage and bone, induction of adhesion molecules, induction of procoagulant activity in vascular endothelial cells, increased adhesion of neutrophils and lymphocytes, and activation of macrophages, neutrophils and vascular endothelial cells. Inflammatory induction that causes tissue damage such as stimulating the release of platelet-activating factor from endothelial cells causes the following effects.

[0006] TNFα is associated with infections, immune disorders, tumor pathologies, autoimmune pathologies, and graft-versus-host disease states. The association between TNFα and cancer and infectious pathologies is often related to the catabolic state of the host. Cancer patients are usually troubled by weight loss associated with loss of appetite.

[0007] The significant debilitation associated with cancer and other diseases is known as "cachexia." Cachexia is characterized by progressive weight loss, loss of appetite, and persistent decline in fat-free mass in response to the growth of malignant tumors. The cachectic state contributes to the morbidity and mortality of many cancers. There is evidence that TNFα is involved in cachexia in cancer, infectious pathologies, and other catabolic states.

[0008] TNFα is thought to play a central role in gram-negative sepsis and endotoxin shock, including fever, malaise, loss of appetite, and cachexia. Endotoxin strongly activates monocyte / macrophage production and the secretion of TNFα and other cytokines. TNFα and other monocyte-derived cytokines mediate the metabolic and neurohormonal responses to endotoxin. Administration of endotoxin to human volunteers results in an acute disease accompanied by symptoms such as fever, tachycardia, increased metabolic rate, and stress hormone release, similar to those of influenza. The circulation of TNFα increases in patients suffering from gram-negative sepsis. Therefore, TNFα is associated with inflammatory diseases, autoimmune diseases, viral, bacterial, and parasitic infections malignant tumors, and / or neurodegenerative diseases, and is associated with rheumatoid arthritis and Crohn's

[0009] disease. disease. It is a target useful for specific biological therapies in diseases such as leprosy. The beneficial effects in an open-label trial using a chimeric monoclonal antibody (cA2) against TNFα have been reported, including suppression of inflammation and good retreatment after recurrence in rheumatoid arthritis and Crohn's disease. Beneficial results in a randomized double-blind placebo-controlled trial using cA2 have also been reported, including suppression of inflammation in rheumatoid arthritis.

[0010] Other researchers have described mAbs specific for recombinant human TNF with neutralizing activity in vitro. Some of these mAbs have been used to map the epitopes of human TNF and to develop enzyme immunoassays and to assist in the purification of recombinant TNF. However, these studies do not provide a basis for generating TNF-neutralizing antibodies that can be used for in vivo diagnostic or therapeutic applications in humans due to immunogenicity, low specificity, and / or pharmaceutical inappropriateness.

[0011] Neutralizing antisera or mAbs against TNF have been shown to suppress harmful physiological changes and prevent death after a lethal challenge in experimental endotoxemia and bacteremia in mammals other than humans. This effect has been shown, for example, in rodent lethality assays and primate pathology models.

[0012] The putative receptor-binding sites of hTNF have been disclosed, and the receptor-binding sites of TNFα consisting of amino acids 11-13, 37 -42, 49-57, and 155-157 of TNF have been disclosed.

[0013] Non-human mammals, chimeras, polyclonal (e.g., antiserum), and / or monoclonal Rue antibodies (Mabs) and fragments (e.g., proteolytic digestion or production of their fusion proteins substances) are, in some cases, promising therapeutic agents being investigated for the treatment of certain diseases. However, such antibodies or fragments, when administered to humans, may induce an immune response resulting in immune complex-mediated clearance of the antibody or fragment from the bloodstream, rendering repeated dosing inappropriate for treatment, thereby reducing the therapeutic benefit to the patient and limiting readministration of the antibody or fragment. For example, repeated dosing of antibodies or fragments containing non-human moieties can result in serum sickness and / or anaphylaxis. To avoid these and other problems, as is well known in the art, many approaches have been taken to reduce the immunogenicity of such antibodies and their portions, including chimerization and humanization. However, these and other approaches still result in antibodies or fragments that have some immunogenicity, low affinity, low binding activity, or are associated with problems in cell culture, scale-up, production, and / or low yields. Thus, such antibodies or fragments may not be ideally suited for manufacture or use as therapeutic proteins possibly. Therefore, there was a need to overcome another of these problems with anti-TNF antibodies or fragments and to provide improvements to known antibodies or their fragments. From this need, SIMPONI® (golimumab), a fully human monoclonal anti-TNF antibody, was developed

[0014]

[0015] SUMMARY OF THE INVENTION MEANS FOR SOLVING THE PROBLEM

[0015] In certain embodiments, the present invention provides a method for treating a patient's active ankylosing spondylitis The method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder such that a statistically significant improvement in disease activity is confirmed by week 16 of treatment compared to patients treated with placebo and the improvement is maintained or improved through week 52 of treatment, and the disease activity is determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the a Short-Form-36 Physical Component Summary (SF-36 PCS), the mean change from baseline in the a Short-Form-36 Mental Component Summary (SF-36 MCS), the mean change from baseline in a Mixed-effect Repeated Measures statistical model (MOS-SS), and the mean change from baseline in the EuroQol-5D visual analog scale (EQ-VAS).

[0016] In certain embodiments, the present invention provides a method for treating a patient's active ankylosing spondylitis The method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof​​​​​ including administration to a patient, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, wherein the patient is responsive to treatment and has a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved through week 52 of treatment, and the statistically significant improvement in disease activity by week 16 of treatment is an average change from baseline in AS QoL of -5.4 ± 5.0 standard deviations (SD), an average change from baseline in SF-36 PCS of 8.5 ± 7.5 SD, an average change from baseline in SF-36 MCS of 6.5 ± 9.1 SD, an average change from baseline in MOS-SS of 6.6 ± 7.2 SD, and an average change from baseline in EQ-VAS of 20.3 ± 24 .6 SD, and is selected from the group consisting of In certain embodiments, the present invention provides a method for treating active ankylosing spondylitis in a patient

[0017] the method including administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, wherein the patient is responsive to treatment and has a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved through week 52 of treatment, and the disease activity is an average change from baseline in the quality of life in ankylosing spondylitis (AS QoL) and an average change from baseline in 36-item Short-Form Physical Component Summary (SF-36 PCS) and is selected from the group consisting of and is selected from the group consisting of Mean change, mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and is determined by a response selected from the group consisting of the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and is administered by intravenous infusion (IV) such that the antibody is administered at a dose of 2 mg / kg and administered over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks (q8w) thereafter. In certain embodiments, the present invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder to treatment, having a statistically significant improvement in disease activity by week 16 of treatment as compared to patients treated with placebo,

[0018] the improvement being maintained or improved through week 52 of treatment, and the disease activity being determined by a response selected from the group consisting of mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), mean change from baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS), mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and the composition is administered by intravenous infusion (IV) such that the antibody is administered at a dose of 2 mg / kg and administered over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks (q8w) thereafter. In certain embodiments, the present invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder to treatment, having a statistically significant improvement in disease activity by week 16 of treatment as compared to patients treated with placebo, the improvement being maintained or improved through week 52 of treatment, and the disease activity being determined by a response selected from the group consisting of mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), mean change from baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS), mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and the composition is administered by intravenous infusion (IV) such that the antibody is administered at a dose of 2 mg / kg The body is administered at a dose of 2 mg / kg, at week 0 and week 4, and then every 8 weeks (q8 w), and is administered by intravenous injection (IV) so as to be administered over 30 ± 10 minutes, and this method further comprises administering the composition regardless of the presence or absence of methotrexate (MTX), sulfasalazine (SSZ), or hydroxychloroquine (HCQ). In certain embodiments, the present invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof,

[0019] wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder such that there is a statistically significant improvement in disease activity by week 16 of treatment as compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity is determined by a response selected from the group consisting of the mean change from baseline in the quality of life in ankylosing spondylitis (AS QoL), the mean change from baseline in the 36-item Short-Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), and the method further comprises, before, simultaneously with, or after said administration, a detectable label or reporter, a TNF antagonist, an anti-ly ​​​​Hippiatric drugs, muscle relaxants, anesthetics, non-steroidal anti-inflammatory drugs (NSAIDs), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blockers, antibacterial drugs, anti-psoriatic agents, corticosteroids, anabolic steroids, erythropoietin, immunizing agents, immunoglobulins, immunosuppressive drugs, growth hor mones, hormone replacement drugs, radiopharmaceuticals, antidepressants, antipsychotics, stimulants, asthma drugs, beta-agonist drugs, inhaled steroids, epinephrine or analogs, cytokines, or cytokine antag onists, at least one compound or protein selected from at least one of the above, and further comprising administering at least one composition comprising an effective dose thereof. In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS).

[0020] In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, having a statistically significant improvement in disease activity by week 16 of the treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of the treatment, the disease activity being the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) scale, the mean change from baseline of the 36-item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline of the 36-item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). The response is determined by a response selected from the group consisting of: mean change from baseline

[0021] In certain embodiments, the present invention provides a method for the treatment of active ankylosing spondylitis in a patient. The method further provides a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof. and administering to the patient an antibody comprising the anti-TNF antibody having a polypeptide comprising the amino acid sequence of SEQ ID NO:36. and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is treated with Responders to CI, compared with placebo-treated patients, through week 16 of treatment A statistically significant improvement in disease activity was observed in patients with refractory glaucoma, which persisted through 52 weeks of treatment This statistically significant improvement in disease activity by week 16 of treatment was consistent with AS Mean change from baseline in QoL = -5.4 ± 5.0 standard deviations (SD), SF-3 6 Mean change from baseline in PCS = 8.5 ± 7.5 SD, SF-36 MCS = 1.0 ± 1.0 SD Mean change from baseline = 6.5 ± 9.1 SD, MOS-SS from baseline Mean change = 6.6 ± 7.2 SD, mean change from baseline in EQ-VAS = 20.3 ±24.6SD.

[0022] In certain embodiments, the present invention provides a method for the treatment of active ankylosing spondylitis in a patient. The method further provides a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof. and administering to the patient an antibody comprising the anti-TNF antibody having a polypeptide comprising the amino acid sequence of SEQ ID NO:36. and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is treated with Responders to CI, compared with placebo-treated patients, through week 16 of treatment it is confirmed that there is a statistically significant improvement in disease activity, and the improvement persists until week 52 of treatment is maintained or improved, and the disease activity is based on the mean change from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) mean change from the baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS) mean change from the baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS) mean change from the baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS) and mean change from the baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and is determined by a response selected from the group consisting of the composition is administered by intravenous infusion (IV) such that the anti-TNF antibody or its antigen-binding fragment is administered at a dose of 2 mg / kg over 30 ± 10 minutes at weeks 0 and 4, and then every 8 weeks (q8w) thereafter

[0023] In certain embodiments, the present invention provides a composition for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, and having been confirmed to have a statistically significant improvement in disease activity until week 16 of treatment compared to a patient treated with placebo and the improvement is maintained or improved until week 52 of treatment, and the disease activity is based on the mean change from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) mean change from the baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS) is maintained or improved, and the disease activity is based on the mean change from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL) mean change from the baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS) ​​​​Mean change from baseline, 36-Item Short-Form Mental Component Summary (SF-36 MCS) Mean change from baseline, mean change from baseline of the mixed effects repeated measures statistical model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), determined by a response selected from the group consisting of mean change from baseline, and the method further comprises administering the composition regardless of the presence or absence of methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ).

[0024] In certain embodiments, the present invention provides a composition for use in the treatment of a patient's active ankylosing spondylitis, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder to the treatment and has been confirmed to have a statistically significant improvement in disease activity by week 16 of the treatment compared to patients treated with placebo, and the improvement is maintained or improved until week 52 of the treatment, and the disease activity is determined by a response selected from the group consisting of mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), mean change from baseline of the 36-Item Short-Form Physical Component Summary (SF-36 PCS), mean change from baseline, mean change from baseline of the 36-Item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline, mean change from baseline of the mixed effects repeated measures statistical model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), and the method comprises ​​Before, at the same time as, or after administration, a detectable label or reporter, TNF antagonist drug, antirheumatic drug, muscle relaxant, anesthetic, non-steroidal anti-inflammatory drug (NSAID), analgesic anesthetic, sedative, local anesthetic, neuromuscular blocker, antibacterial agent, anti-psoriatic agent, corticosteroid anabolic steroid, erythropoietin, immunizing agent, immunoglobulin, immunosuppressive agent growth hormone, hormone replacement drug, radiopharmaceutical, antidepressant, antipsychotic, stimulant, asthma drug , β-agonist, inhaled steroid, epinephrine or analog, cytokine, or cytokine at least one compound or polypeptide selected from at least one of the antagonists, and administering to the patient an effective amount of at least one composition comprising the same. In certain embodiments, the present invention provides a method for treating active ankylosing spondylitis in a patient

[0025] which method comprises administering to the patient an anti-TNF antibody or antigen-binding fragment thereof wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37 and the patient is a responder to treatment such that there is a statistically significant improvement in disease activity by week 16 of treatment as compared to patients treated with placebo and the improvement is maintained or improved through week 52 of treatment, and the disease activity is the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short-Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Outcomes in Rheumatology (MOS-SS) repeated measures statistical model, and the Euro and the improvement is maintained or improved through week 52 of treatment, and the disease activity is the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short-Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Outcomes in Rheumatology (MOS-SS) repeated measures statistical model, and the Euro the mean change from baseline in the 36-Item Short-Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Outcomes in Rheumatology (MOS-SS) repeated measures statistical model, and the Euro the mean change from baseline in the Mixed Outcomes in Rheumatology (MOS-SS) repeated measures statistical model, and the Euro Determined by a response selected from a group consisting of the mean change from the baseline of the Qol-5D Visual Analogue Scale (EQ-VAS). It is determined by a response selected from a group consisting of the change.

[0026] In certain embodiments, the present invention provides a method for treating a patient's ankylosing spondylitis. The method includes administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof. The anti-TNF antibody includes a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37. The patient is a responder to the treatment and has a statistically significant improvement in disease activity by week 16 of the treatment compared to patients treated with placebo. The improvement is maintained or improved until week 52 of the treatment, and the statistically significant improvement in disease activity by week 16 of the treatment is from the baseline of AS QoL with a mean change = -5.4 ± 5.0 standard deviation (SD), from the baseline of SF-36 PCS with a mean change = 8.5 ± 7.5 SD, from the baseline of SF-36 MCS with a mean change = 6.5 ± 9.1 SD, from the baseline of MOS-SS with a mean change = 6.6 ± 7. 2 SD, and from the baseline of EQ-VAS with a mean change = 20.3 ± 24.6 SD, and is selected from the group consisting of these. The statistically significant improvement in disease activity by week 16 of the treatment is from the baseline of AS QoL with a mean change = -5.4 ± 5.0 standard deviation (SD), from the baseline of SF-36 PCS with a mean change = 8.5 ± 7.5 SD, from the baseline of SF-36 MCS with a mean change = 6.5 ± 9.1 SD, from the baseline of MOS-SS with a mean change = 6.6 ± 7. 2 SD, and from the baseline of EQ-VAS with a mean change = 20.3 ± 24.6 SD, and is selected from the group consisting of these. 2 SD, and from the baseline of EQ-VAS with a mean change = 20.3 ± 24.6 SD, and is selected from the group consisting of these. 2 SD, and from the baseline of EQ-VAS with a mean change = 20.3 ± 24.6 SD, and is selected from the group consisting of these. 2 SD, and from the baseline of EQ-VAS with a mean change = 20.3 ± 24.6 SD, and is selected from the group consisting of these. It is selected from the group consisting of these.

[0027] In certain embodiments, the present invention provides a method for treating a patient's ankylosing spondylitis. The method includes administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof. The anti-TNF antibody includes a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37. The patient is a responder to the treatment and has a statistically significant improvement in disease activity by week 16 of the treatment compared to patients treated with placebo. The improvement is maintained or improved until week 52 of the treatment, and the statistically significant improvement in disease activity by week 16 of the treatment is from the baseline of AS QoL with a mean change = -5.4 ± 5.0 standard deviation (SD), from the baseline of SF-36 PCS with a mean change = 8.5 ± 7.5 SD, from the baseline of SF-36 MCS with a mean change = 6.5 ± 9.1 SD, from the baseline of MOS-SS with a mean change = 6.6 ± 7. No improvement was confirmed, and the improvement was maintained or improved until week 52 of treatment, and the disease activity was the mean change from baseline in the quality of life in ankylosing spondylitis (AS QoL), 3 the mean change from baseline in the 6-item Short-Form Physical Component Summary (SF-36 PCS), 36 the mean change from baseline in the 36-item Short-Form Mental Component Summary (SF-36 MCS), the mixed-effects mean change from baseline in the repeated measures statistical model (MOS-SS), and the mean change from baseline in the EuroQol -5D Visual Analogue Scale (EQ-VAS), and was determined by the response selected from the group consisting of The anti-TNF antibody or its antigen-binding fragment was administered at a dose of 2 mg / kg, and was administered by intravenous infusion (IV) such that it was administered over 30 ± 10 minutes at weeks 0 and 4, and then every 8 weeks (q8w) thereafter.

[0028] In certain embodiments, the present invention provides a method of treating active ankylosing spondylitis in a patient comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder to the treatment, and has a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, and the improvement is maintained or improved until week 52 of treatment, and the disease activity is the mean change from baseline in the quality of life in ankylosing spondylitis (AS QoL), 3 the mean change from baseline in the 6-item Short-Form Physical Component Summary (SF-36 PCS), 36 ​​The mean change from baseline in the Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), determined by a response selected from the group consisting of, the method further comprising administering the anti-TNF antibody or antigen-binding fragment thereof, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present. In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, and In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, administering, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present, the anti-TNF antibody or antigen-binding fragment thereof.

[0029] In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, administering, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present, the anti-TNF antibody or antigen-binding fragment thereof. In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, administering, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present, the anti-TNF antibody or antigen-binding fragment thereof. In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, administering, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present, the anti-TNF antibody or antigen-binding fragment thereof. In certain embodiments, the invention provides a method for treating a patient's active ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment, as confirmed by a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, the disease activity being determined by a response selected from the group consisting of the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-Item Short Form Physical Component Summary (SF-36 PCS), the mean change from baseline in the 36-Item Short Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), the method further comprising, prior to said administration, administering, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present, the anti-TNF antibody or antigen-binding fragment thereof. Simultaneously with, or after administration, a detectable label or reporter, a TNF antagonist, an antirheumatic drug, a muscle relaxant, an anesthetic, a non-steroidal anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antibacterial agent, an antipsoriatic agent, a corticosteroid, an anabolic steroid, erythropoietin, an immunizing agent, an immunoglobulin, an immunosuppressive agent, a growth hormone, a hormone replacement agent, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma drug, a β-agonist, an inhaled steroid, epinephrine or an analog, a cytokine, or a cytokine antagonist, and administering at least one composition comprising an effective amount of at least one compound or protein selected from at least one of these. In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS)

[0030] In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, the patient being a responder to the treatment and having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-36 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the Mixed Effects Repeated Measures Statistical Model (MOS-SS) The average change from the baseline, and the average change from the baseline of the EuroQol-5D Visual Analogue Scale is determined by a response selected from the group consisting of the average change from the baseline thereof.

[0031] In certain embodiments, the present invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of a patient's ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37 and the patient is a responder to the treatment and has been confirmed to have a statistically significant improvement in disease activity by week 16 of the treatment as compared to a patient treated with placebo, and the improvement is maintained or improved until week 52 of the treatment, and the said statistically significant improvement in disease activity by week 16 of the treatment is selected from the group consisting of an average change from the baseline of AS QoL = -5.4 ± 5.0 standard deviation (SD), an average change from the baseline of SF-36 PCS = 8.5 ± 7.5 SD , an average change from the baseline of SF-36 MCS = 6.5 ± 9.1 SD, an average change from the baseline of MOS-S S = 6.6 ± 7.2 SD, and an average change from the baseline of EQ-VAS = 20.3 ± 24.6 SD. In certain embodiments, the present invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of a patient's ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37

[0032] In certain embodiments, the present invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of a patient's ankylosing spondylitis, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37 ​​ comprising, and the patient is a responder to treatment, compared to a patient treated with placebo and there is a statistically significant improvement in disease activity by week 16 of treatment, and the improvement is maintained or improved until week 52 of treatment, and the disease activity is the mean change from baseline in ankylosing spondylitis quality of life (AS QoL), the mean change from baseline in the 36-item Short-Form Physical Component Summary (SF-3 6 PCS), the mean change from baseline in the 36-item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS) and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), and is determined by a response selected from the group consisting of the mean change from baseline and the anti-TNF antibody or antigen-binding fragment thereof is administered by intravenous infusion ( IV) such that the anti-TNF antibody or antigen-binding fragment thereof is administered at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks (q8w) thereafter. IV).

[0033] In certain embodiments, the invention provides an anti-TNF antibody or antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37 and the patient is a responder to treatment, compared to a patient treated with placebo and there is a statistically significant improvement in disease activity by week 16 of treatment, and the improvement is maintained or improved until week 52 of treatment, and the disease activity is the mean change from baseline in ankylosing spondylitis quality of life is maintained or improved until week 52 of treatment, and the disease activity is the mean change from baseline in the quality of life in ankylosing spondylitis (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-3 6 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the mean change from baseline in the mixed effects repeated measures statistical model (MOS-SS) , and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-VAS), and is determined by a response selected from the group consisting of the method further comprising administering the anti-TNF antibody or an antigen-binding fragment thereof, whether or not methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) is present.

[0034] In certain embodiments, the invention provides an anti-TNF antibody or an antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient, the method comprising administering to the patient an anti-TNF antibody or an antigen-binding fragment thereof, the anti-TNF antibody comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37 , the patient being a responder to the treatment, having been confirmed to have a statistically significant improvement in disease activity by week 16 of treatment compared to a patient treated with placebo, the improvement being maintained or improved until week 52 of treatment, and the disease activity being the mean change from baseline in the quality of life in ankylosing spondylitis (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-3 6 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 (AS QoL), the mean change from baseline in the 36-item Short Form Physical Health Summary (SF-3 6 PCS), the mean change from baseline in the 36-item Short Form Mental Health Summary (SF-36 MCS), the mean change from baseline in the mixed effects repeated measures statistical model (MOS-SS) The mean change from the baseline, and the mean change from the baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), as determined by a response selected from the group consisting of determined, the method comprising, prior to, simultaneously with, or after said administration, administering at least one composition comprising an effective amount of at least one compound or protein selected from a detectable label or reporter, TNF antagonist, anti-rheumatic drug, muscle relaxant, narcotic, non-steroidal anti-inflammatory drug ( NSAID), analgesic, anesthetic, sedative, local anesthetic, neuromuscular blocker, antibacterial, anti-psoriatic agent, corticosteroid, anabolic steroid, erythropoietin, immunizing agent, immunoglobulin agent, immunosuppressive agent, growth hormone, hormone replacement agent, radiopharmaceutical, antidepressant, antipsychotic agent, stimulant, asthma drug, beta agonist, inhaled steroid, epinephrine or analog, cytokine, or cytokine antagonist. Further included.

[0035] In another embodiment, the present invention provides a method for treating active ankylosing spondylitis in a patient, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is responsive to the treatment, showing a statistically significant improvement in disease activity by week 16 of treatment as compared to patients treated with placebo, the improvement being maintained or improved until about week 52 of treatment, and the disease activity being the mean change from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from the baseline of the 36-Item Short-Form Physical Component Summary (SF-36 PCS) from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), the mean change from the baseline of the 36-Item Short-Form Physical Component Summary (SF-36 PCS) from the baseline of the 36-Item Short-Form Physical Component Summary (SF-36 PCS) Mean change, mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS). It is determined by a response selected from the group consisting of mean changes from baseline.

[0036] In another embodiment, the present invention provides a method for treating a patient with active ankylosing spondylitis, the method comprising administering to the patient a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof, wherein the anti-TNF antibody comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, and the patient is a responder such that there is a statistically significant improvement in disease activity by week 16 of treatment compared to patients treated with placebo, and the improvement is maintained or improved through week 52 of treatment, and the disease activity is determined by a response comprising one or more of the mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), mean change from baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS), mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS), mean change from baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS), and mean change from baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS).

[0037] In another embodiment, the present invention provides a mean change from baseline of the Ankylosing Spondylitis Quality of Life (AS QoL), mean change from baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS), or mean change from baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS). the mean change of these, from the baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS) the mean change, the mean change from the baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS) , and the mean change from the baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS), or one or more of the equivalents thereof, as determined by a response that includes is made

[0038] In another embodiment, the present invention provides a method for treating active ankylosing spondylitis in a patient that includes administering to the patient a composition comprising means for contacting TNF , wherein the patient is a responder to the treatment and has been confirmed to have an improvement in disease activity by the 16th week of treatment, compared to patients treated with placebo , and the improvement is maintained or improved until about the 52nd week of treatment, and the disease activity is determined by a response that includes one or more of the mean change from the baseline of the Ankylosing Spondylitis Quality of Life (AS QoL )), the mean change from the baseline of the 36-item Short-Form Physical Component Summary (SF-36 PCS) , the mean change from the baseline of the 36-item Short-Form Mental Component Summary (SF-36 MCS) , the mean change from the baseline of the Mixed Effects Repeated Measures Statistical Model (MOS-SS) , and the mean change from the baseline of the EuroQol-5D Visual Analogue Scale (EQ-VAS ). )

[0039] In another embodiment, the present invention provides a method for treating active ankylosing spondylitis in a patient that includes administering to the patient a pharmaceutical composition comprising means for contacting TNF , wherein the patient is a responder to the treatment and, compared to patients treated with placebo such that improvement in disease activity is confirmed by week 16 of treatment, and the improvement is maintained or improved until week 52 of treatment, and the disease activity is determined by a response including one or more of the mean change from baseline in the quality of life in ankylosing spondylitis (AS Q oL), the mean change from baseline in the 36-item Short-Form Physical Component Summary (SF-36 PCS ), the mean change from baseline in the 36-item Short-Form Mental Component Summary (SF-36 MCS), the mean change from baseline in the Mixed Effects Repeated Measures Statistical Model (MOS-SS ), and the mean change from baseline in the EuroQol-5D Visual Analogue Scale (EQ-V AS). In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis, wherein the anti-TNF antibody is administered by intravenous (IV) infusion, and a patient treated with the anti-TNF antibody achieves an ASDAS inactive disease (<1.3) at week 4 or

[0040] week 2 of treatment. In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis, wherein the anti-TNF antibody is administered by intravenous (IV) infusion at week 0 and week 4, and then every 8 weeks (q8w) thereafter for 30 ± 10 minutes at a dose of 2 mg / kg, and a patient treated with the anti-TNF antibody achieves an ASDAS inactive disease (<1.3) at week 4 or week 2 of treatment.

[0041] In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis, wherein the anti-TNF antibody is administered by intravenous (IV) infusion at week 0 and week 4, and then every 8 weeks (q8w) thereafter for 30 ± 10 minutes at a dose of 2 mg / kg, and a patient treated with the anti-TNF antibody achieves an ASDAS inactive disease (<1.3) at week 4 or week 2 of treatment, and ​​At least one isolated patient will achieve ASDAS inactive disease (<1.3) at the second week of treatment. The present invention provides mammalian anti-TNF antibodies.

[0042] The present invention relates to a heavy chain (HC) comprising SEQ ID NO: 36 for use in the treatment of active ankylosing spondylitis. ) and at least one isolated mammalian anti-T having a light chain (LC) comprising SEQ ID NO:37. NF antibody, the antibody is an anti-NF antibody that is effective against methotrexate (MTX), sulfasalazine (SSZ ) or hydroxychloroquine (HCQ), The antibody is administered by intravenous (IV) infusion, and patients treated with the anti-TNF antibody are Achieve ASDAS inactive disease (<1.3) at least at week 4 of treatment or week 2 of treatment An isolated mammalian anti-TNF antibody is provided.

[0043] In certain embodiments, the present invention provides a compound, At least one having a heavy chain (HC) comprising sequence number 36 and a light chain (LC) comprising sequence number 37 An isolated mammalian anti-TNF antibody and at least one pharma- ceutically acceptable carrier or and a diluent, wherein the composition is administered by IV infusion, and Treated patients had ASDAS inactive disease (<1.3) at week 4 of treatment or week 2 of treatment The present invention provides a composition that achieves the above object.

[0044] In certain embodiments, the present invention provides a compound, At least one having a heavy chain (HC) comprising sequence number 36 and a light chain (LC) comprising sequence number 37 An isolated mammalian anti-TNF antibody and at least one pharma- ceutically acceptable carrier or A composition comprising a diluent, wherein the composition is administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks thereafter, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. administered at and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. Provided is a composition.

[0045] In certain embodiments, the invention is for use in the treatment of ankylosing spondylitis, at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, and at least one pharmaceutically acceptable carrier or diluent, wherein the composition is administered with or without including methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition is administered IV, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. diluent, wherein the composition is administered with or without including methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition is administered IV, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. diluent, wherein the composition is administered with or without including methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition is administered IV, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. diluent, wherein the composition is administered with or without including methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition is administered IV, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. diluent, wherein the composition is administered with or without including methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition is administered IV, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. Provided is a composition.

[0046] In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. ). Provided is a method.

[0047] In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is ankylosing spondylitis, the method comprising administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition is administered by IV infusion, and the patient treated with the composition achieves ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4 and then every 8 weeks (q8w) thereafter, and the patient being treated with the composition achieving an ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. provided. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered with or without comprising methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition being administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4 and then every 8 weeks (q8w) thereafter, and the patient being treated with the composition achieving an ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. provided. In certain embodiments, the invention is a method for treating a TNF-related condition,

[0048] wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion, and the composition wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered with or without comprising methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition being administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4 and then every 8 weeks (q8w) thereafter, and the patient being treated with the composition achieving an ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. provided. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion, and the composition wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered with or without comprising methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition being administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4 and then every 8 weeks (q8w) thereafter, and the patient being treated with the composition achieving an ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. provided. In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion, and the composition

[0049] wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion, and the composition wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered with or without comprising methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), the composition being administered by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4 and then every 8 weeks (q8w) thereafter, and the patient being treated with the composition achieving an ASDAS-inactive disease (<1.3) at week 4 of treatment or week 2 of treatment. provided. wherein the TNF-related condition is active ankylosing spondylitis and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, the composition being administered by IV infusion, and the composition Patients treated with [the treatment] achieve an ASDAS inactive disease (<1.3 ) at week 4 of treatment or week 2 of treatment, and the method comprises, before, at the same time as, or after said administration, a detectable label or a porter, a TNF antagonist, an anti-rheumatic drug, a muscle relaxant, a narcotic, a non-steroidal anti-inflammatory drug (N SAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antibacterial drug, an anti-psoriatic drug, a corticosteroid, an anabolic steroid, erythropoietin, an immunizing agent, an immunoglobulin a, an immunosuppressive drug, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic drug, a stimulant, an asthma drug, a beta-agonist, an inhaled steroid, epinephrine or an analog, a cytokine, or a cytokine antagonist, and further comprises administering at least one composition comprising an effective amount of at least one compound or protein selected from at least one of the foregoing. A method is provided.

[0050] In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis, wherein the anti-TNF antibody is administered by intravenous (IV) infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody have a Bath Ankylosing Spondylitis Functional Index (BASF I) = -2.4 ± 2.1 standard deviations (SD), a Bath Ankylosing Spondylitis Metrology Index (Ba th Ankylosing Spondylitis Metrology Index, BASMI) = -0.4 ± 0.6 SD, and ​​36-Item Short-Form Health Survey Physical Component Summary (SF-36 PCS) = 8.5 ± 7.5 SD, 36-Item Short-Form Health Survey Mental Component Summary (SF-36 MCS) = 6.5 ± 9.1 SD, and Ankylosing Spondylitis Quality of Life (ASQoL) = -5.4 ± 5.0 SD, and achieve an average change from baseline in one or more criteria selected from the group consisting of: Provide at least one isolated mammalian anti-TNF antibody. In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, wherein the anti-TNF antibody is administered at 0 and 4 weeks, and then every 8 weeks (q8w) thereafter, by intravenous (IV) infusion at a dose of 2 mg / kg over 30 ± 10 minutes, and at 16 weeks of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD. Provide at least one isolated mammalian anti-TNF antibody.

[0051]

[0052] ​​​​​​​​​​​​​​In certain embodiments, the invention is an array for use in the treatment of ankylosing spondylitis having at least one heavy chain (HC) comprising SEQ ID NO: 36 and at least one light chain (LC) comprising SEQ ID NO: 37 which is at least one isolated mammalian anti-TNF antibody, wherein the anti-TNF antibody is administered with or without methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), and is administered by intravenous (IV) infusion at a dose of 2 mg / kg, and wherein, at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD, thereby providing at least one isolated mammalian anti-TNF antibody

[0053] In certain embodiments, the invention is a composition comprising at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, and at least one pharmaceutically acceptable carrier or diluent, wherein the composition is administered by IV infusion and wherein, at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD, thereby providing a composition

[0054] ​​​​​​​​​ In certain embodiments, the present invention provides a composition for use in the treatment of ankylosing spondylitis, the composition comprising at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, and at least one pharmaceutically acceptable carrier or diluent, wherein the composition is administered by intravenous infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks (q8w) thereafter, and wherein, at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD.

[0055] In certain embodiments, the present invention provides a composition for use in the treatment of ankylosing spondylitis, the composition comprising at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, and at least one pharmaceutically acceptable carrier or diluent, wherein the composition is administered with or without methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), and wherein the composition is administered by intravenous infusion, and wherein, at week 16 of treatment, patients treated with the anti-TNF antibody have BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 SD ​​​​​​​、and achieving an average change from baseline in one or more criteria selected from the group consisting of ASQoL = -5.4 ± 5.0 SD Provide a composition that achieves an average change from baseline in one or more criteria selected from the group consisting of ASQoL = -5.4 ± 5.0 SD

[0056] In certain embodiments, the present invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, wherein the composition is administered by IV infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, B ASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-3 6 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD Provide a method that achieves an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, B ASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-3 6 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD

[0057] In certain embodiments, the present invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37, wherein the composition is administered at week 0 and week 4, and then every 8 weeks (q8w) thereafter, by IV infusion at a dose of 2 mg / kg over 30 ± 10 minutes and at week 16 of treatment, patients treated with the anti-TNF antibody have BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7. 5 SD, SF-36 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD achieve an average change from a baseline in one or more criteria selected from the group consisting of SD A method is provided that comprises

[0058] In certain embodiments, the invention is a method for treating a TNF-related condition, comprising wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 wherein the composition is administered with or without comprising methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ) wherein the composition is administered by IV infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 S D, SF-36 PCS = 8.5 ± 7.5 SD, SF-36 MCS = 6.5 ± 9.1 S D, and ASQoL = -5.4 ± 5.0 SD A method is provided that comprises In certain embodiments, the invention is a method for treating a TNF-related condition, comprising wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 wherein the composition is administered by IV infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, B

[0059] In certain embodiments, the invention is a method for treating a TNF-related condition, comprising wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises administering a composition comprising an isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 wherein the composition is administered by IV infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, BASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-3 wherein the composition is administered by IV infusion, and at week 16 of treatment, patients treated with the anti-TNF antibody achieve an average change from baseline in one or more criteria selected from the group consisting of BASFI = -2.4 ± 2.1 SD, B ASMI = -0.4 ± 0.6 SD, SF-36 PCS = 8.5 ± 7.5 SD, SF-3 6 MCS = 6.5 ± 9.1 SD, and ASQoL = -5.4 ± 5.0 SD a group consisting of 6 MCS = 6.5 ± 9.1 SD and ASQoL = -5.4 ± 5.0 SD, or achieve an average change from baseline in one or more criteria selected from , prior to, simultaneously with, or after said administration, a detectable label or reporter, a TNF antagonist drug, antirheumatic drug, muscle relaxant, narcotic, non-steroidal anti-inflammatory drug (NSAID), analgesic anesthetic, sedative, local anesthetic, neuromuscular blocker, antibacterial, anti-psoriatic, corticosteroid anabolic steroid, erythropoietin, immunizing agent, immunoglobulin, immunosuppressive agent growth hormone, hormone replacement, radiopharmaceutical, antidepressant, antipsychotic, stimulant, asthma drug , beta agonist, inhaled steroid, epinephrine or analog, cytokine, or a cytokine antagonist, and further comprising administering at least one composition comprising at least one compound or protein selected from at least one of these .

[0060] In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis wherein said anti-TNF antibody is administered by intravenous (IV) infusion and wherein at least 65% of the patients treated achieve ASAS20 at week 16 of treatment . In certain embodiments, the invention provides at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis

[0061] wherein said anti-TNF antibody is administered by intravenous (IV) infusion and wherein at least 1 one of said isolated mammalian anti-TNF antibodies administered more than 65% of the patients who received the treatment achieved ASAS20 at week 16 of the treatment with a treatment difference of at least 45% (improvement compared to placebo). There is provided at least one isolated mammalian anti-TNF antibody.

[0062] In certain embodiments, the invention relates to at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis. administered by intravenous (IV) infusion at a dose of 2 mg / kg over 30 ± 10 minutes at week 0 and week 4, and then every 8 weeks (q8w) thereafter, and more than 65% of the patients who received the treatment achieved ASAS20 at week 16 of the treatment. There is provided at least one isolated mammalian anti-TNF antibody.

[0063] In certain embodiments, the invention relates to at least one isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 36 and a light chain (LC) comprising SEQ ID NO: 37 for use in the treatment of active ankylosing spondylitis. administered by intravenous (IV) infusion, with or without methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), and more than 65% of the patients who received the treatment achieved ASAS20 at week 16 of the treatment. There is provided at least one isolated mammalian anti-TNF antibody.

[0064] In certain embodiments, the invention provides a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises a heavy chain comprising SEQ ID NO: 36. An isolated mammalian anti-TNF antibody having a heavy chain (HC) comprising a lock (HC) and a light chain (LC) comprising SEQ ID NO: 37 administering a composition comprising, wherein the composition is administered by IV infusion and is undergoing treatment such that at least 65% of the patients achieve ASAS20 at week 16 of treatment, a method is provided .

[0065] In certain embodiments, the invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises a heavy chain comprising SEQ ID NO: 36 administering a composition comprising an isolated mammalian anti-TNF antibody having a light chain (LC) comprising SEQ ID NO: 37 wherein the composition is administered by IV infusion and at least 65% of the patients undergoing treatment achieve ASAS20 at week 16 of treatment with a treatment difference of at least 45% (improvement compared to placebo ), a method is provided.

[0066] In certain embodiments, the invention provides a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis and the method comprises a heavy chain (HC ) and an isolated mammalian anti-TNF antibody having a light chain (LC) comprising SEQ ID NO: 37, the composition is administered at 0 and 4 weeks and then every 8 weeks thereafter ( q8w) at a dose of 2 mg / kg over 30 ± 10 minutes via IV infusion and at least 65% of the patients undergoing treatment achieve ASAS20 at week 16 of treatment.

[0067] The invention is a method for treating a TNF-related condition, wherein the TNF-related condition is active ankylosing spondylitis, and the method comprises a heavy chain (HC) comprising SEQ ID NO: 36 and a SEQ ID NO: 3 Administering a composition comprising an isolated mammalian anti-TNF antibody having a light chain (LC) comprising 7 therewith, and the composition is administered by IV infusion, with or without methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine (HCQ), and in which more than 65% of the patients being treated achieve ASAS20 by week 16 of treatment provided a method.

Brief Description of the Drawings

[0068]

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Mode for Carrying Out the Invention

[0069] The present invention relates to all of the heavy chain variable CDR regions of SEQ ID NOs: 1, 2, and 3, and / or all of the light chain variable CDR regions of SEQ ID NOs: 4, 5, and 6, and includes an isolated recombinant and / or synthetic anti-TNF human, primate, rodent, mammalian, chimeric, humanized or CDR-grafted antibodies, and anti-TNF anti-idiotype antibodies thereto, and provides a composition and a coding nucleic acid molecule comprising at least one polynucleotide encoding at least one anti-TNF antibody or anti-idiotype antibody. The present invention further includes, but is not limited to, diagnostic and therapeutic compositions, methods, and devices, and methods for the production and use of such nucleic acids and antibodies, and anti-idiotype antibodies. ​​

[0070] As used herein, "anti-tumor necrosis factor α antibody", "anti-TNF antibody", "anti-TNF antibody portion" or "anti-TNF antibody fragment", and / or "anti-TNF antibody variant", etc., are at least one complementarity-determining region (CDR) of the heavy or light chain, or its ligand-binding portion, the variable region of the heavy or light chain, the constant region of the heavy or light chain, the framework region, or any portion thereof, or at least one portion of a TNF receptor or binding protein, etc., but not limited thereto, and include molecules containing at least a part of an immunoglobulin molecule, and include any protein or peptide-containing molecule. Such an antibody may, if desired, further affect a specific ligand, but is not limited thereto. Such an antibody can, in vitro, in situ, and / or in vivo, regulate, reduce, increase, antagonize, activate, relieve, alleviate, block, inhibit, suppress, and / or interfere with at least one TNF activity or binding or TNF receptor activity or binding. As a non-limiting example, a preferred anti-TNF antibody, a specified portion or variant of the present invention can bind to at least one TNF or a specified portion, variant or domain thereof. A preferred anti-TNF antibody, a specified portion or variant can also, if desired, affect at least one of TNF activities or functions such as RNA, DNA, or protein synthesis, TNF release, TNF receptor signaling, membrane TNF cleavage, TNF activity, TNF production, and / or synthesis, but not limited thereto. The term "antibody" further includes antibodies, their digestion fragments, is intended to include antibody mimetics, or portions of antibodies that mimic the structure and / or function of an antibody, or specific fragments or portions thereof, including single-chain antibodies and their fragments. Functional fragments include antigen-binding fragments that bind to mammalian TNF. For example, Fab (e.g., by papain digestion), Fab’ (e.g., by pepsin digestion and partial reduction), and F(ab’)2 (e.g., by pepsin digestion), fac b (e.g., by plasmin digestion), pFc’ (e.g., by pepsin or plasmin digestion), Fd (e.g., by pepsin digestion, partial reduction, and reassembly), Fv, or sc Fv (e.g., by molecular biological techniques) fragments, but are not limited thereto. Antibody fragments that can bind to TNF or portions thereof are encompassed by the present invention (see, e.g., Colligan, Immunology supra).

[0071] Such fragments can be produced by enzymatic cleavage, synthesis, or recombinant techniques, as known in the art and / or as described herein. Antibodies can be produced in various truncated forms using an antibody gene in which one or more stop codons have been introduced upstream of the natural stop site. For example, combinations of genes encoding the F(ab’)2 heavy chain portion can be designed to include DNA sequences encoding the CH1 domain and / or hinge region of the heavy chain. Various portions of the antibody can be chemically conjugated by conventional techniques, or prepared as a continuous protein without interruption using genetic engineering techniques.

[0072] As used herein, the term “human antibody” refers to substantially all portions of the protein (e.g., For example, CDR, framework, C L , C H domain (e.g., C H 1, C H 2, C H 3 ), hinge (V L , V H )) refers to an antibody that is substantially non-immunogenic in humans with only minor sequence changes or mutations. Similarly, antibodies designated for primates (such as monkeys, baboons, chimpanzees, etc.), rodents (such as mice, rats, rabbits, guinea pigs, hamsters, etc.), and other mammalian animals refer to antibodies specific to such species, subgenera, genera, subfamilies, and families. Furthermore, chimeric antibodies include any combination of the above. Such changes or mutations may, optionally and preferably, retain or reduce immunogenicity in humans or other species compared to the unmodified antibody. Thus, human antibodies are different from chimeric or humanized antibodies. Hu man antibodies are produced by animals other than humans, or prokaryotic or eukaryotic cells that can express functionally rearranged human immunoglobulin (e.g., heavy chain and / or light chain) genes It is pointed out that it can be. Moreover, when a human antibody is a single-chain antibody, it may contain a linker peptide not found in natural human antibodies. For example, Fv may contain a linker peptide such as 2 to about 8 glycine or other amino acid residues that connect the variable region of the heavy chain and the variable region of the light chain . Such a linker peptide is considered to be of human origin.

[0073] Also, bispecific, heterospecific, heteroconjugate, or similar antibodies that are monoclonal, preferably human or humanized antibodies having binding specificities for at least two different antigens may be used. In this case, one of the binding specificities is at least one TNF ta ​ one directed against a protein and the other against any other antigen. Methods for producing bispecific antibodies are known in the art. Conventionally, recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 30 5:537 (1983)). Due to the random combination of immunoglobulin heavy and light chains , these hybridomas (quadromas) produce a possible mixture of 10 different antibody molecules , only one of which has the correct bispecific structure. Purification of the correct molecules ( usually performed by affinity chromatography procedures) is quite cumbersome and the yield of the product is low. Similar procedures are described, for example, in WO 93 / 08829, US Pat. No. 6 ,210,668, 6,193,967, 6,132,992, 6, 106,833, 6,060,285, 6,037,453, 6, 10,902, 5,989,530, 5,959,084, 5,95 9,083, 5,932,448, 5,833,985, 5,821 ,333, 5,807,706, 5,643,759, 5,601, 819, 5,582,996, 5,496,549, 4,676,9 80, WO 91 / 00360, 92 / 00373, EP 0308 9, Traunecker et al., EMBO J. 10:3655 (1991 ), Suresh et al., Methods in Enzymology 12 Disclosed in 1:210(1986), each of which is hereby incorporated by reference in its entirety. into this specification.

[0074] Anti-TNF antibodies (also referred to as TNF antibodies) useful in the methods and compositions of the present invention may optionally be characterized by high affinity binding to TNF and, optionally and preferably, low toxicity. Specifically, individual components such as variable regions, constant regions, and frameworks may individually and / or collectively, optionally and preferably, have low immunogenicity and the antibodies, specifically identified fragments, or variants of the present invention are useful in the present invention. Antibodies that can be used in the present invention may optionally be characterized based on their ability to treat patients over a long period of time with a measurable alleviation of symptoms and low and / or acceptable toxicity. Low or acceptable immunogenicity, and / or high affinity, and other suitable properties can contribute to the treatment results obtained. "Low immunogenicity" as used herein is defined as an increase in HAHA, HACA or HAMA responses in less than about 75%, or preferably less than about 50%, of the patients being treated, and / or low titers (less than about 300, preferably less than about 100 as measured by a double antigen enzyme immunoassay) in the patients being treated (Elliott et al., Lancet 3 44:1125-1127(1994), which is hereby incorporated by reference in its entirety). ) (Elliott et al., Lancet 3 44:1125-1127(1994), which is hereby incorporated by reference in its entirety).

[0075] Utility: The isolated nucleic acids of the present invention can be measured in or act on cells, tissues, organs or animals (including mammals and humans) to treat immune disorders or diseases, circulatory disorders or diseases, infectious disorders or diseases, , at least one TNF state selected from but not limited to at least one of malignant and / or neurological disorders or diseases can be used to diagnose, monitor, regulate, treat, alleviate, prevent the occurrence of, or reduce the symptoms of, at least one TNF state, and can be used to generate at least one anti-TNF antibody or a specified variant thereof.

[0076] Such a method may include administering to a cell, tissue, organ, animal or patient in need of such regulation, treatment, alleviation, prevention, or reduction of symptoms, effects, or mechanisms, an effective amount of a composition or pharmaceutical composition comprising at least one anti-TNF antibody. The effective amount can be determined using known methods described herein or known in the relevant art, and can be, for example, a single (e.g., bolus), multiple, or continuous dose of about 0.001 - 500 mg / kg of body weight, or a single, multiple, or continuous dose that achieves a serum concentration of 0.01 - 5000 μg / mL, or any effective range or value therein. Cited references. All publications or patents cited in this specification are hereby incorporated by reference in their entirety into this specification, showing the state of the art at the time of the present invention, and / or providing an explanation and enabling use of the present invention. Publications refer to any scientific publication, patent gazette, or any other information available in any media format, including those recorded in electronic or printed form. The following references are hereby incorporated by reference in their entirety into this specification: Ausubel, et al., ed., , Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, NY (1987 - 2001), Sa , Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, NY (1987 - 2001), Sa mbrook, et al., Molecular Cloning: A Labora tory Manual, 2nd Edition, Cold Spring Harb or, NY(1989), Harlow and Lane, antibodies, a Laboratory Manual, Cold Spring Harbor, NY (1989), Colligan, et al., eds., Current Prot ocols in Immunology, John Wiley & Sons, Inc. , NY(1994 - 2001), Colligan et al., Current P rotocols in Protein Science, John Wiley & S ons, NY, NY, (1997 - 2001).

[0077] Antibodies of the present invention: all of the heavy chain variable CDR regions of SEQ ID NOs: 1, 2, and 3, and / or all of the light chain variable CDR regions of SEQ ID NOs: 4, 5, and 6, at least one anti-TN F antibody of the present invention can, if desired, be produced by a cell line, a mixed cell line, an immortalized cell or a clonal population of immortalized cells, as is well known in the art. For example, Ausubel , et al., ed., Current Protocols in Molecul ar Biology, John Wiley & Sons, Inc., NY, NY(19 87 - 2001), Sambrook, et al., Molecular Cloni ng: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, NY(1989), Harlow and Lane, a ntibodies, a Laboratory Manual, Cold Sprin g Harbor, NY (1989), Colligan, et al., eds., C current Protocols in Immunology,John Wile y&Sons, Inc., NY (1994-2001), Colligan et al. .,Current Protocols in Protein Science,J See Wiley & Sons, NY, NY, (1997-2001). Each is incorporated herein by reference in its entirety.

[0078] Human antibodies specific for human TNF protein or fragments thereof are isolated and / or Suitable molecules such as TNF proteins or portions thereof (including synthetic molecules such as synthetic peptides) Other specific or general mammalian antibodies may be raised against any suitable immunogenic antigen. Preparation of immunogenic antigens and generation of monoclonal antibodies can be accomplished by any suitable technique. This can be done using techniques.

[0079] In one approach, hybridomas are generated using a suitable immortalized cell line (e.g., Sp2 / 0 , Sp2 / 0-AG14, NSO, NS1, NS2, AE-1, L.5, >243, P3 X63Ag8.653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 S A5, U937, MLA 144, ACT IV, MOLT4, DA-1, JURKAT , WEHI, K-562, COS, RAJI, NIH 3T3, HL-60, MLA 1 Myeloma cells, such as, but not limited to, 44, NAMAIWA, and NEURO 2A strains, or heteromylomas, fusion products thereof, or any derived therefrom. by fusing with cells or fused cells, or any other suitable cell line known in the art For example, see www.atcc.org, www.life tech.com, etc. Antibody-producing cells such as isolated or cloned spleen, peripheral blood, lymph, tonsils, or other immune or B cell-containing cells, but not limited to these, or recombinant or endogenous, viruses, bacteria, algae, prokaryotes, amphibians, insects, reptiles, fish, mammals, rodents, horses, sheep, goats, sheep, primates class, eukaryotes, genomic DNA, cDNA, rDNA, mitochondrial DNA or RN A, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single-stranded, double-stranded or triple-stranded, hybridized, etc., or combinations thereof, as either endogenous or heterologous nucleic acids, having any other cell that expresses heavy or light chain constant or variable or framework or CDR sequences. For example, see Ausubel, supra, and Chapter 2 of Colligan, Immunology. Both references are hereby incorporated by reference in their entirety.

[0080] Antibody-producing cells can also be obtained from the peripheral blood of humans or other suitable animals immunized with the antigen of interest, or preferably from the spleen or lymph nodes. Using any other suitable host cell, heterologous nucleic acids or endogenous nucleic acids encoding the antibodies, specific fragments or variants thereof of the present invention can also be expressed. Fused cells (hybridomas) or recombinant cells are isolated using selective culture conditions or other suitable known methods, and limiting dilution or It can be cloned by cell sorting or other known methods. Having the desired specificity Cells that produce antibodies with the desired specificity can be selected by a suitable assay (e.g., ELISA). It can be done.

[0081] Methods for selecting recombinant antibodies from peptide or protein libraries are mentioned, but this is not limited to them. Other methods suitable for producing or isolating antibodies having the required specificity can be used (e.g., display libraries such as bacteriophage, ribosome, oligonucleotide, RNA, cDNA, etc., but not limited to these. For example, available from Cambridge antibody Technologies, Cambridge shire, UK, MorphoSys, Martinsreid / Planegg, DE, Biovation, Aberdeen, Scotland, UK, BioInvent, Lund, Sweden, Dyax Corp., Enzon, Affymax / Biosite, Xoma, Berkeley, CA, Ixsys.). For example, European Patent No. 368,684, International Application PCT / GB91 / 01134, International Application PCT / GB92 / 01755, International Application PCT / GB92 / 002240, International Application PCT / GB92 / 00883, International Application PCT / GB93 / 00605, US Patent Application No. 08 / 350260 (5 / 12 / 94), International Application P CT / GB94 / 01422, International Application PCT / GB94 / 02662, International Application P CT / GB97 / 01835, (CAT / MRC), International Publication No. 90 / 14443, International Publication No. 90 / 14424, International Publication No. 90 / 14430, International Application PCT / US CT / GB94 / 02662, International Application PCT / GB97 / 01835, (CAT / MRC), International Publication No. 90 / 14443, International Publication No. 90 / 14424, International Publication No. 90 / 14430, International Application PCT / US No. 94 / 1234, International Publication No. WO 92 / 18619, International Publication No. WO 96 / 07754 (Sc ripps), European Patent No. 614 989 (MorphoSys), International Publication No. WO 95 / 16027 (BioInvent), International Publication No. WO 88 / 06630, International Publication No. WO 90 / 3809 (Dyax), US Patent No. 4,704,692 (Enzon), International Application PCT / US91 / 02989 (Affymax), International Publication No. WO 89 / 06283, Eu ropean Patent No. 371998, European Patent No. 550400, (Xoma), European Patent No. 229 046, International Application PCT / US91 / 07149 (Ixsys), or a stochastically generated peptide or protein - US Patent Nos. 5723323, 576319 2, 5814476, 5817483, 5824514, 59 76862, International Publication No. WO 86 / 05803, European Patent No. 590689 (Ixsys , now Applied Molecular Evolution (AME), each of which is , incorporated herein by reference in its entirety), or known in the art and / or described herein, and that can generate a repertoire of human antibodies and that depends on immunization of transgenic animals (e.g., SCID mice, Ngu yen et al., Microbiol. Immunol. 41:901 - 907( 1997), Sandhu et al., Crit. Rev. Biotechnol. 16:95 - 118(1996), Eren et al., Immunol. 93:1 54 - 161(1998) (each of which is incorporated herein by reference in its entirety). Such techniques include ribosome display (Hanes et al., Proc. Natl. Acad Sci. . Sci. USA, 94:4937 - 4942 (May 1997), Hanes et al., Proc. Natl. Acad. Sci. USA, 95:14130 - 141 35 (Nov. 1998)), single - cell antibody production techniques (e.g., selected lymphocyte antibody method ( “SLAM”) (U.S. Patent No. 5,627,052, Wen et al., J. Imm unol. 17:887 - 892 (1987), Babcock et al., Pro c. Natl. Acad. Sci. USA 93:7843 - 7848 (1996)), germ microwells and flow cytometry (Powell et al., Biotechnol. 8:333 - 337 (1990), One Cell Syst ems, Cambridge, MA, Gray et al., J. Imm. Meth. 182:155 - 163 (1995), Kenny et al., Bio / Techn ol. 13:787 - 790 (1995), B - cell selections (Steenbakkers et al., Molec. Biol. Reports 19:125 - 134 (199 4), Jonak et al., Progress Biotech, Vol. 5, I n Vitro Immunization in Hybridoma Techno logy, Borrebaeck, ed., Elsevier Science Pub lishers B.V., Amsterdam, Netherlands (1988) ) are mentioned, but not limited to these.

[0082] Methods for engineering or humanizing non - human antibodies or human antibodies can be used similarly, and the It is well known in the art. Generally, humanized or engineered antibodies have one or more amino acid residues from non-human sources such as, for example, mice, rats, rabbits, non-human primates, or other mammals, but are not limited thereto. These human amino acid residues are often referred to as "import" residues and are typically taken from the "import" variable regions, constant regions, or other domains of known human sequences.

[0083]

[0083] Known human Ig sequences are disclosed in numerous publications and websites, for example, www.ncbi.nlm.nih.gov / entrez / query.fcgi; www.atcc.org / phage / hdb.html; www.sciquest.com / ; www.abcam.com / ; www.antibodyresource.com / onlinecomp.htm l; www.public.iastate.edu / ~pedro / research_ tools.html; www.mgen.uni-heidelberg.de / SD / IT / IT.htm l; www.whfreeman.com / immunology / CH05 / kuby0 5.htm; www.library.thinkquest.org / 12429 / Immun e / Antibody.html; www.hhmi.org / grants / lectures / 1996 / vlab; www.path.cam.ac.uk / ~mrc7 / mikeimages.htm l; www.antibodyresource.com / ; www.mcb.harvard.edu / BioLinks / Immunology .html. www.immunologylink.com / ; www.pathbox.wustl.edu / ~hcenter / index.ht ml; www.biotech.ufl.edu / ~hcl / ; www.pebio.com / pa / 340913 / 340913.html; www.nal.usda.gov / awic / pubs / antibody / ; www.m.ehime-u.ac.jp / ~yasuhito / Elisa.htm l; www.biodesign.com / table.asp; www.icnet.uk / axp / facs / davies / links.html ; www.biotech.ufl.edu / ~fccl / protocol.html ; www.isac-net.org / sites_geo.html; www.aximt1.imt.uni-marburg.de / ~rek / AEPS tart.html; www.baserv.uci.kun.nl / ~jraats / links1.ht ml; www.recab.uni-hd.de / immuno.bme.nwu.edu / ; www.mrc-cpe.cam.ac.uk / imt-doc / public / IN TRO.html; www.ibt.unam.mx / vir / V_mice.html;imgt.cn usc.fr:8104 / 、 www.biochem.ucl.ac.uk / ~martin / abs / ind ex.html; antibody.bath.ac.uk / 、 www.abgen.cvm.tamu.edu / lab / www.abgen.html; www.unizh.ch / ~honegger / AHOseminar / Slide 01.html; www.cryst.bbk.ac.uk / ~ubcg07s / ; www.nimr.mrc.ac.uk / CC / ccaewg / ccaewg.htm ; www.path.cam.ac.uk / ~mrc7 / humanisation / T AHHP.html; www.ibt.unam.mx / vir / structure / stat_aim. html; www.biosci.missouri.edu / smithgp / index.h tml; www.cryst.bioc.cam.ac.uk / ~fmolina / Web-p ages / Pept / spottech.html; www.jerini.de / frproducts.html; www.patents.ibm.com / ibm.html.Kabat et a l., Sequences of Proteins of Immunological Interest, U.S. Dept. Health(1983) exists, and each is incorporated herein by reference in its entirety.

[0084] Such imported sequences are used to reduce immunogenicity or in the art ​As is known in the art, it can be used to reduce, enhance or modify binding, affinity, association rate constant, dissociation rate constant, binding activity, specificity , half-life, or any other suitable property. Generally, some or all of the non-human or human CDR sequences are maintained while the non-human sequences of the variable and constant regions are replaced with human or other amino acids. Antibodies can also be humanized while retaining high affinity for the antigen and other desirable biological properties, if desired. To achieve this purpose, if desired, the humanized antibody can be prepared by an analysis process of the parental sequence and various conceptual humanized products using three-dimensional models of the parental sequence and the humanized sequence . Three-dimensional immunoglobulin models are generally available and well known to those skilled in the art. Computer programs are available that illustrate and display the likely three-dimensional conformations of selected immunoglobulin sequence candidates. By examining these displays, it is possible to analyze the likely high function of residues in the function of the immunoglobulin sequence candidate, i.e., the residues that affect the antigen-binding ability of the immunoglobulin candidate . In this way, FR residues can be selected and combined from the consensus sequence and the import sequence so that desirable antibody properties, such as increased affinity for the target antigen(s), are achieved. Generally, CDR residues directly and almost substantially affect antigen binding. The humanization or engineering of the antibodies of the present invention is based on Winter (Jones s et al., Nature 321:522 (1986), Riechmann et al., Nature 332:323 (1988), Verhoeyen et al., Science 239:1534 (1988)), Sims et al. . Three-dimensional immunoglobulin models are generally available and well known to those skilled in the art. Computer programs are available that illustrate and display the likely three-dimensional conformations of selected immunoglobulin sequence candidates. By examining these displays, it is possible to analyze the likely high function of residues in the function of the immunoglobulin sequence candidate, i.e., the residues that affect the antigen-binding ability of the immunoglobulin candidate . In this way, FR residues can be selected and combined from the consensus sequence and the import sequence so that desirable antibody properties, such as increased affinity for the target antigen(s), are achieved. Generally, CDR residues directly and almost substantially affect antigen binding. The humanization or engineering of the antibodies of the present invention is based on Winter (Jones s et al., Nature 321:522 (1986), Riechmann et al., Nature 332:323 (1988), Verhoeyen et al., Science 239:1534 (1988)), Sims et al. . Thus, FR residues can be selected and combined from the consensus sequence and the import sequence so that desirable antibody properties, such as increased affinity for the target antigen(s), are achieved. Generally, CDR residues directly and almost substantially affect antigen binding. The humanization or engineering of the antibodies of the present invention is based on Winter (Jones s et al., Nature 321:522 (1986), Riechmann et al., Nature 332:323 (1988), Verhoeyen et al., Science 239:1534 (1988)), Sims et al. s et al., Nature 321:522(1986), Riechmann et al., Nature 332:323(1988), Verhoeyen et al., Science 239:1534(1988)), Sims et al. , J. Immunol. 151:2296 (1993), Chothia and Le sk, J. Mol. Biol. 196:901 (1987), Carter et al ., Proc. Natl. Acad. Sci. U.S.A. 89:4285 (1992) 、Presta et al., J. Immunol. 151:2623 (1993)、 U.S. Patent No. 5,723,323, 5,976,862, 5,824,514, 5,8 17,483, 5,814,476, 5,763,192, 5,723,323, 5,766,886, 5,714,352, 6,204,023, 6,180 370, 5,693,762, 5,530,101, 5,585,089, 5,225,539, 4,816,567, International Application PCT / :US98 / 16280 、US96 / 18978, US91 / 09630, US91 / 05939, US9 4 / 01234, GB89 / 01334, GB91 / 01134, GB92 / 01 755, International Publication Nos. 90 / 14443, 90 / 14424, 90 / 144 30, European Patent No. 229,246 (each incorporated herein by reference in its entirety, including the references cited therein), etc., but not limited thereto, and can be performed using any known method.

[0085] The anti-TNF antibody can also, if desired, be generated by immunizing transgenic animals (e.g., mice, rats, hamsters, non-human primates, etc.) that can generate a repertoire of human antibodies as described herein and / or known in the art. It is also possible. Cells producing human anti-TNF antibodies can be isolated from such animals and immortalized using suitable methods such as those described herein.

[0086] Transgenic mice capable of producing a repertoire of human antibodies that bind to human antigens can be created by known methods (e.g., but not limited to, U.S. Patent Nos. 5,770,428, 5,569, 825, 5,545,806, 5,625,126, 5,625,8 25, 5,633,425, 5,661,016, and 5,789, 650, issued to Lonberg et al.; International Publication Nos. 98 / 50433, 98 / 24893, 98 / 2488 4, 97 / 13852, 94 / 25585, 96 / 34096 issued to Lonberg et al.; International Publication Nos. issued to Kucherlapate et al.; European Patent Nos. 0463151 (B1), 0710719 (A1) issued to Kucherla pate et al.; U.S. Patent No. 5,5 45,807 issued to Surani et al.; International Publication No. 90 / 04036, European Patent No. 0438474 (B1) issued to Brugg emann et al.; European Patent No. 0 814259 (A2) issued to Lonberg et al.; British Patent No. 2272440 (A) issued to Lonberg et al.; Lonberg et al., Nature 368:856 - 859 (1994); T aylor et al., Int. Immunol. 6(4) 579 - 591 (19 ​​​​94), Green et al, Nature Genetics 7:13-21( 1994), Mendez et al., Nature Genetics 15:1 46-156(1997), Taylor et al., Nucleic Acids Research 20(23):6287-6295(1992), Tuaillo n et al., Proc Natl Acad Sci USA 90(8)372 0-3724(1993), Lonberg et al., Int Rev Immu nol 13(1):65-93(1995), and Fishwald et al., Nat Biotechnol 14(7):845-851(1996), which are incorporated herein by reference in their entirety). Generally, these mice contain at least one transgene comprising DNA derived from at least one human immunoglobulin locus that is functionally rearranged or capable of undergoing functional rearrangement. The endogenous immunoglobulin loci of such mice can be disrupted or deleted to eliminate the ability of the mice to produce antibodies encoded by the endogenous genes. Screening for antibodies specific for a similar protein or fragment can be successfully accomplished using a peptide display library. This method involves screening a large number of samples of peptides for individual members with the desired function or structure. Antibody screening of peptide display libraries is well known in the art. The length of the displayed peptide sequences is 3-5

[0087] ​​​​​​​​000 or more amino acids, frequently 5 to 100 amino acids in length, often about 8 to 25 amino acids in length. In addition to direct chemical synthesis methods for creating peptide libraries, several recombinant DNA methods have also been described. One type involves the display of peptide sequences on the surface of bacteriophages or cells. Each bacteriophage or cell contains a nucleotide sequence encoding a specific displayed peptide sequence. Such methods are described in International Publication Nos. WO 91 / 17271, WO 91 / 18980, WO 91 / 19818, and WO 93 / 08278. Other systems for creating peptide libraries have aspects of both chemical synthesis and recombinant methods in vitro. See International Publication Nos. WO 92 / 05258, WO 92 / 14843, and WO 96 / 19256. Also see U.S. Pat. Nos. 5,658,754 and 5,64 3,768. Peptide display libraries, vectors, and screening kits are commercially available from suppliers such as Invitrogen (Carlsbad, CA) and Cambri dge antibody Technologies (Cambridgeshire , UK). For example, U.S. Pat. Nos. 4,704,692, 4,939,666, 4,946,778, 5,260,20 3, 5,455,030, 5,518,889, 5,534,621, 5,6 56,730, 5,763,733, 5,767,260, 5,856,456, assigned to Enzon; U.S. Pat. Nos. 5,223,409, 5,403,484, 5,57 1,698, 5,837,500, assigned to Dyax; U.S. Pat. No. 5,427,9 assigned to Affymax; ​ No. 08, the same as U.S. Patent No. 5,580,717, Cambridge antibody Techno logies, U.S. Patent No. 5,885,793 assigned to Genentech, U.S. Patent No. 5,750,373 assigned to Xoma, U.S. Patent No. 5,618,920 assigned to Xoma, the same as No. 5,595,898, the same as No. 5,576,195, the same as No. 5,698,435, the same as No. 5,693,49 3, the same as No. 5,698,417, see Colligan above, Ausubel above, or Sambrook above referred to above. Each of the above patents and publications is hereby incorporated by reference in its entirety into this specification.

[0088] The antibodies of the present invention can also be prepared using at least one nucleic acid encoding an anti-TNF antibody to provide transgenic animals or mammals such as goats, cows, horses, sheep, etc. that produce such antibodies in milk. Such animals can be prepared using known methods . For example, but not limited to, U.S. Patent Nos. 5,827,69 0, 5,849,992, 4,873,316, 5,849,992 No., 5,994,616, 5,565,362, 5,304,489 etc. (each of which is hereby incorporated by reference in its entirety into this specification).

[0089] The antibodies of the present invention can be further prepared using at least one anti-TNF antibody-encoding nucleic acid to provide transgenic plants and cultured plant cells (e.g., tobacco and corn, but not limited thereto) that produce such antibodies, the identified parts or variants in plant parts or cells cultured therefrom . Non-limiting examples As such, for example, using an inducible promoter, transgenic tobacco leaves expressing recombinant proteins have been successfully used to provide large amounts of recombinant proteins. For example, see Cramer et al., Curr. Top. Microbiol. Immunol. 240: 95-118 (1999) and the references cited therein. Also, transgenic maize has been used to express mammalian proteins having biological activities equivalent to those of proteins produced in other recombinant systems or purified from natural sources at commercial production levels. For example, see Hood et al., Adv. Exp. Med. Biol. 464: 127-147 (1999) and the references cited therein. Antibodies have also been produced in large quantities from transgenic plant seeds containing antibody fragments such as single-chain antibodies (scFv) including tobacco seeds and potato tubers. For example, see Conrad et al., Plant Mol. Biol. 38: 101-109 (1998) and the references cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants by known methods. For example, see Fischer et al., Biotechnol. Appl. Biochem. 30: 99-108 (Oct., 1999), Ma et al., Trends Biotechnol. 13: 522-7 (1995), Ma et al., Plant Physiol. 109: 341-6 (1995), Whitelam et al., Biochem. Soc. Trans. 22: 940-944 (1994), and the references cited therein. Also, although not limited, generally antibodies have been successfully used to provide large amounts of recombinant proteins. For example, see Cr amer et al., Curr. Top. Microbiol. Immunol. 24 0: 95-118 (1999) and the references cited therein. Also, transgenic maize has been used to express mammalian proteins having biological activities equivalent to those of proteins produced in other recombinant systems or purified from natural sources at commercial production levels. For example, see Hood et al., Adv . Exp. Med. Biol. 464: 127-147 (1999) and the references cited therein. Antibodies have also been produced in large amounts from transgenic plant seeds containing antibody fragments such as single-chain antibodies (s cFv) including tobacco seeds and potato tubers. For example, see Conrad et al., Plant Mol. Biol. 38: 101 -109 (1998) and the references cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants by known methods. For example, see Fischer et al., Biotechnol. Appl. Bio . Exp. Med. Biol. 464: 127-147 (1999) and the references cited therein. Antibodies have also been produced in large amounts from transgenic plant seeds containing antibody fragments such as single-chain antibodies (s cFv) including tobacco seeds and potato tubers. For example, see Conrad et al., Plant Mol. Biol. 38: 101 -109 (1998) and the references cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants by known methods. For example, see Fischer et al., Biotechnol. Appl. Biochem. 30: 99-108 (Oct., 1999), Ma et al., Tren ds Biotechnol. 13: 522-7 (1995), Ma et al., P lant Physiol. 109: 341-6 (1995), Whitelam et . Exp. Med. Biol. 464: 127-147 (1999) and the references cited therein. Antibodies have also been produced in large amounts from transgenic plant seeds containing antibody fragments such as single-chain antibodies (s cFv) including tobacco seeds and potato tubers. For example, see Conrad et al., Plant Mol. Biol. 38: 101 -109 (1998) and the references cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants by known methods. For example, see Fischer et al., Biotechnol. Appl. Biochem. 30: 99-108 (Oct., 1999), Ma et al., Tren ds Biotechnol. 13: 522-7 (1995), Ma et al., P lant Physiol. 109: 341-6 (1995), Whitelam et . Exp. Med. Biol. 464: 127-147 (1999) and the references cited therein. Antibodies have also been produced in large amounts from transgenic plant seeds containing antibody fragments such as single-chain antibodies (s cFv) including tobacco seeds and potato tubers. For example, see Conrad et al., Plant Mol. Biol. 38: 101-109 (1998) and the references cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants by known methods. For example, see Fischer et al., Biotechnol. Appl. Biochem. 30: 99-108 (Oct., 1999), Ma et al., Trends Biotechnol. 13: 522-7 (1995), Ma et al., Plant Physiol. 109: 341-6 (1995), Whitelam et al., Biochem. Soc. Trans. 22: 940-944 (1994), and the references cited therein. Also, although not limited, generally antibodies See also for plant expression. Each of the above documents is incorporated herein by reference in its entirety.

[0090] The antibodies of the present invention can bind to human TNF with a wide range of affinities (K D ). In a preferred embodiment, at least one human mAb of the present invention can bind to human TN F with high affinity. For example, the human mAb can bind human TNF at about 10 - 7 M or less, such as 0.1 - 9.9 (or any range or value therein) X10 -7 , 1 0 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 or any range or value therein (but not limited to these) of K D .

[0091] The affinity or binding activity of an antibody for an antigen can be determined experimentally using any suitable method. (See, for example, Berzofsky, et al., "Antibody- Antigen Interactions," In Fundamental Imm unology, Paul, W.E., Ed., Raven Press: New Yo rk, NY (1984), Kuby, Janis Immunology, W.H. Fr eeman and Company: New York, NY (1992), and the methods described in this specification). The affinity measured for a particular antibody-antigen interaction can vary when measured under different conditions (e.g., salt concentration, pH). Thus . See also the methods described in this specification). The affinity measured for a particular antibody-antigen interaction can vary when measured under different conditions (e.g., salt concentration, pH). Thus the affinity can be different when measured under different conditions (e.g., salt concentration, pH). Therefore ​That is, the measurement of affinity and other antigen-binding parameters (e.g., K D、 K a 、K d ) is preferably performed using standard solutions of the antibody and antigen, and a standard buffer such as the buffer described herein.

[0092] Nucleic acid molecule. A nucleotide sequence encoding at least 70-100% of at least one contiguous amino acid of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, a specified fragment, variant or consensus sequence thereof, or using the information provided herein such as a deposited vector containing at least one of these sequences, all of the heavy-chain variable CDR regions of SEQ ID NO: 1, 2 and 3 and / or all of the light-chain variable CDR regions of SEQ ID NO: 4, 5 and 6 The nucleic acid molecule of the present invention encoding at least one anti-TNF antibody can be obtained using the methods described herein or known in the art.

[0093] The nucleic acid molecule of the present invention may be in the form of mRNA, hnRNA, tRNA or any other form of RNA such as , or in the form of DNA including but not limited to cDNA and genomic DNA obtained by cloning or synthetically produced, or any combination thereof. The DNA may be triple-stranded, double-stranded or single-stranded, or any combination thereof. Any part of at least one strand of DNA or RNA may be a coding strand also known as a sense strand, or a non-coding strand called an antisense strand.

[0094] The isolated nucleic acid molecule of the present invention may optionally have one or more introns in an open reading frame ​​​​​-coding frame (ORF), for example, but not limited to, at least one heavy chain (e.g., SEQ ID NOs: 1-3) or light chain (e.g., SEQ ID NOs: 4-6) CDR1, C DR2, and / or at least one specific part of at least one CDR such as CDR3, a nucleic acid molecule, an anti-TNF antibody or a coding sequence of a variable region (e.g., SEQ ID NOs: 7, 8), and a nucleic acid molecule that is substantially different from the above nucleic acid molecule, but due to the degeneracy of the genetic code, encodes at least one anti-TNF antibody described herein and / or known in the art. A nucleic acid molecule may be included. Of course, the genetic code is well known in the art. Therefore, it will be routine for those skilled in the art to generate such degenerate nucleic acid variants that encode a specific anti-TNF antibody of the present invention. For example, see Ausubel et al. above. Such nucleic acid variants are included in the present invention. Non-limiting examples of the isolated nucleic acid molecules of the present invention include, respectively, HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 , LC CDR3, and corresponding non-limiting examples of nucleic acids encoding the HC variable region and LC variable region of SEQ ID NOs: 10, 11, 12, 13, 14, 15.

[0095] As shown herein, the nucleic acid molecules of the present invention containing nucleic acids encoding anti-TNF antibodies encode the amino acid sequence of an antibody fragment by itself, a sequence encoding the full length of an antibody or a part of an antibody , a coding sequence of an antibody, fragment or part, and additional sequences, for example, at least one intron, etc., with or without the aforementioned additional coding sequences, non- 5' and 3' sequences of the code, for example, splicing and polyadenylation signals (e.g., , ribosome binding and stability of mRNA) in transcription, mRNA processing Additional non-coding sequences including, but not limited to, transcribed untranslated sequences that play a role Also included are at least one signal leader or coding sequence of a fusion peptide, additional Amino acids, for example, additional coding sequences encoding amino acids that provide additional functions Can be mentioned, but are not limited thereto. Thus, the sequence encoding the antibody can be fused to a marker Sequence, for example, the marker sequence is a sequence encoding a peptide that facilitates the purification of an antibody fragment Or an antibody containing a portion thereof.

[0096] Polynucleotides that selectively hybridize to the polynucleotides described herein. The present invention provides an isolated nucleic acid that hybridizes to the polynucleotides disclosed herein under selective hybridization Conditions. Thus, the polynucleotides of this embodiment Can be used to isolate, detect, and / or quantify nucleic acids containing such polynucleotides For example, the polynucleotides of the present invention can be used to identify, isolate, or amplify partial-length or full-length clones in a deposited Library. In some embodiments, the polynucleotide is an isolated or otherwise Complementary genomic or cDNA sequence to the cDNA of a human or mammalian nucleic acid library. cDNA sequence.

[0097] Preferably, the cDNA library is at least 80% of the full-length sequence, preferably complete At least 85% or 90% of the long array, and more preferably at least 95 % of the full-length array. This cDNA library can be normalized to increase the expression level of rare sequences. Low or medium stringency hybridization conditions are typical, but not limited to, using sequences with low sequence identity to the complementary sequence. For sequences with higher identity, medium and high stringency conditions can optionally be used. Low stringency conditions allow selective hybridization of sequences with about 70% sequence identity and can be used to identify orthologous or paralogous sequences.

[0098] Optionally, the polynucleotides of the present invention will encode at least a portion of an antibody encoded by the polynucleotides described herein. The polynucleotides of the present invention include nucleic acid sequences that are available for selective hybridization to polynucleotides encoding the antibodies of the present invention. See, for example, Ausubel, supra, and Colligan, supra. Each is incorporated herein by reference in its entirety.

[0099] Construction of nucleic acids. The isolated nucleic acids of the present invention can be made using (a) recombinant methods, (b) synthetic techniques, (c) purification techniques, or combinations thereof, as is well known in the art.

[0100] In addition to the polynucleotides of the present invention, the nucleic acids can conveniently contain sequences. For example, a multiple cloning site containing one or more endonuclease restriction sites can be included in the nucleic acid. ​​​​​​​​​​The nucleic acid can be inserted into the nucleic acid to aid in the isolation of the polynucleotide. can be inserted to aid in the isolation of the translated polynucleotide of the present invention. For example, a hexahistidine marker sequence provides a convenient means for purifying the proteins of the invention. The nucleic acid of the invention (excluding the coding sequence) can be optionally prepared as a polynucleotide of the invention. The present invention is directed to a vector, adapter or linker for the cloning and / or expression of a gene.

[0101] Additional sequences may be added to such cloning and / or expression sequences to facilitate cloning and and / or optimizing their function in expression, aiding in the isolation of polynucleotides. It may be possible to improve the transfer of polynucleotides into cells. The use of vectors, adapters, and linkers is well known in the art. (See, for example, Ausubel, supra, or Sambrook, supra).

[0102] Recombinant methods for constructing nucleic acids. RNA, cDNA, genomic DNA, or any of these. The isolated nucleic acid compositions of the present invention, such as any combination of clones, may be any number of clones known to those of skill in the art. In some embodiments, the present invention can be obtained from biological sources using extraction procedures. An oligonucleotide that selectively hybridizes to a polynucleotide under stringent conditions. A oligonucleotide probe is used to identify a desired sequence in a cDNA or genomic DNA library. Isolation of RNA and construction of cDNA and genomic libraries are well within the skill of the art. (See, e.g., Ausubel, supra, or Sambrook, supra. (This refers to.

[0103] Nucleic Acid Screening and Isolation Methods. A probe based on the nucleotide sequence is used to screen a cDNA or genomic library. Probes can be used to probe genomic or cDNA sequences. By hybridization, homologous genes in the same or different organisms can be isolated. If so, use varying degrees of hybridization stringency in the assay. and either the hybridization or the washing medium can be made stringent. It will be clear that the hybridization conditions can be made more stringent. The greater the degree of complementarity between the probe and target when duplex formation occurs, the greater the degree of complementarity between the probe and target. The degree of stringency can be determined by a variety of factors, such as temperature, ionic strength, pH, and formamide. and the presence of such partially denaturing solvents. The stringency of hybridization can be, for example, within the range of 0% to 50% hormonal concentration. This can be successfully altered by manipulating the amide concentration to change the polarity of the reaction solution. The degree of complementarity (sequence identity) required for possible binding depends on the hybridization medium. and / or the stringency of the washing medium. The degree of complementarity is optimally 1 00%, or 70-100%, or any range or value therein. Small differences in sequence in the probes and primers may affect hybridization and / or It is understood that this can be compensated for by reducing the stringency of the wash medium. It should be understood.

[0104] Methods for amplifying RNA or DNA are well known in the art and are described herein. Based on the teachings and guidance, it can be used in accordance with the present invention without undue experimentation.

[0105] Known methods of DNA or RNA amplification include polymerase chain reaction (PCR) and related amplification processes (e.g., U.S. Patent Nos. 4,683,195; 4,683,202; 4,800,159; 4,965,188 to Mullis et al., U.S. Patent Nos. 4,795,699 and 4,921,794 to Tabor et al., U.S. Patent No. 5,142,033 to Innis, U.S. Patent No. 5,122,464 to Wilson et al., U.S. Patent No. 5,091,310 to Innis, U.S. Patent No. 5,066,584 to Gyllensten et al., U.S. Patent No. 4,889,818 to Gelfand et al., U.S. Patent No. 4,994,370 to Silver et al., U.S. Patent No. 4,766,067 to Biswas, U.S. Patent No. 4,656,134 to Ringold (see these patents), and RNA-mediated amplification (Malek et al., U.S. Patent No. 5,130,238, trade name NASBA) using antisense RNA as a template for double-stranded DNA synthesis for the target sequence, are mentioned but not limited to these (the entire contents of these documents are incorporated herein by reference). (See, for example, Ausubel supra, or Sambrook supra.) For example, the polymerase chain reaction (PCR) technique can be used to amplify the polynucleotides of the present invention and the sequences of related genes directly from genomic DNA or cDNA libraries. PCR and other in vitro amplification methods can also, for example, amplify the protein to be expressed.

[0106] ​​​​cloning a nucleic acid sequence encoding the desired mRNA; detecting the presence of the desired mRNA in a sample; To generate nucleic acids to be used as probes for nucleic acid sequencing or for other purposes. It will be useful in guiding those skilled in the art through in vitro amplification methods. Examples of suitable techniques include Berger, supra; Sambrook, supra; and Ausub, supra. el, and Mullis et al., U.S. Pat. No. 4,683,202 (1987), and In nis, et al., PCR Protocols A Guide to Meth ods and Applications,Eds.,Academic Press Inc, San Diego, CA (1990). Commercially available kits are known in the art. See, for example, the T 4 gene 32 protein (Boehringer Mannheim) was used to identify long P The yield of the CR product can be improved.

[0107] Synthetic Methods for Constructing Nucleic Acids. The isolated nucleic acids of the present invention can be prepared by direct chemical synthesis using known methods. (See, for example, Ausubel et al., supra). Generally, the amplification is performed by hybridization with a complementary sequence or by using a single strand as a template. It is a single-stranded oligonucleotide that can be converted into double-stranded DNA by polymerization with the DNA polymerase used in the Those skilled in the art will appreciate that chemical synthesis of DNA produces oligonucleotides of about 100 or more bases. Although the sequence may be limited, longer sequences can be obtained by ligation of shorter sequences. You will realize that you can.

[0108] Recombinant expression cassette. The present invention further provides a recombinant expression cassette comprising the nucleic acid of the present invention. Using the nucleic acid sequence of the present invention, for example, the cDNA or genomic sequence encoding the antibody of the present invention, a recombinant expression cassette that can be introduced into at least one desired host cell can be constructed. The recombinant expression cassette typically comprises a polynucleotide of the present invention operably linked to a transcriptional start regulatory sequence that directs the transcription of the polynucleotide in the intended host cell. Both heterologous and non-heterologous (i.e., endogenous) promoters can be used to direct the expression of the nucleic acid of the present invention.

[0109] In some embodiments, an isolated nucleic acid that functions as a promoter, enhancer, or other element is introduced at an appropriate position (upstream, downstream, or within an intron) of a non-heterologous form of the polynucleotide of the present invention to upregulate or downregulate the expression of the polynucleotide of the present invention. For example, the endogenous promoter can be altered by mutation, deletion, and / or substitution, either in vivo or in vitro.

[0110] Vectors and host cells. The present invention also relates to vectors comprising the isolated nucleic acid molecules of the present invention, host cells genetically engineered with recombinant vectors, and the production of at least one anti-TNF antibody by recombinant techniques well known in the art. For example, reference is made to Sambrook et al. and Ausubel et al. supra, each of which is incorporated herein by reference in its entirety.

[0111] The polynucleotide can optionally be ligated to a vector containing a selectable marker for host growth. Generally, plasmid vectors are transfected by calcium phosphate precipitation. ​ It is introduced into a precipitate like an object or into a complex with a charged lipid. When the vector is a virus , it can be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.

[0112] The DNA insert should be operably linked to an appropriate promoter. The expression construct further includes a ribosome binding site for translation within the transcription start site, the transcription termination site, and the transcribed region. The coding portion of the mature transcript expressed by the construct preferably includes the translation start site first, and a stop codon (e.g., UAA, UGA or UAG) is appropriately located at the end of the translated mRNA. UAA and UAG are preferred for expression in mammalian or eukaryotic cells .

[0113] The expression vector preferably includes at least one selectable marker, but this is optional . Such markers include, for example, methotrexate (MTX ) for eukaryotic cell culture, dihydrofolate reductase (DHFR, U.S. Patent Nos. 4,399,216, 4, 634,665, 4,656,134, 4,956,288, 5,1 49,636, 5,179,017, ampicillin, neomycin (G418) , mycophenolic acid or glutamine synthetase (GS, U.S. Patent Nos. 5,122,46 4, 5,770,359, 5,827,739) resistance genes, and tetracycline or ampicillin resistance genes for culture in Escherichia coli and other bacteria or prokaryotes, but are not limited thereto (the above patents are hereby incorporated by reference in their entirety into this specification ). (to be incorporated into). Suitable culture media and conditions for the above host cells are known in the art. Suitable vectors will be readily apparent to the person skilled in the art. Introduction of the vector construct into the host cells can be accomplished by calcium phosphate transfection, transfection mediated by DEAE-dextran, transfection mediated by cationic lipids, electroporation, transduction, infection or other known methods. Such methods are described in the above Sambrook, Chapters 1-4 and 16-18, the above Ausubel, Chapters 1, 9, 13, 15, 16, etc., in the art. For such methods, see the above Sambrook, Chapters 1-4 and Chapters 16-18, the above Ausubel, Chapters 1, 9, 13, 15, 16, etc., described in the art.

[0114] At least one antibody of the present invention can be expressed in a modified form such as a fusion protein, and may contain not only a secretion signal but also additional heterologous functional regions. For example, a region of additional amino acids, especially charged amino acids, can be added to the N-terminus of the antibody to improve stability and persistence in the host cells during purification or subsequent processing and storage. Also, a peptide moiety can be added to the antibody of the present invention to facilitate purification. Such regions can be removed before the final preparation of the antibody or at least one of its fragments. Such methods are described in the above Sambrook, Chapters 17.29-17.42 and 18.1-18.74, the above A usubel, Chapters 16, 17 and 18, etc., in many standard laboratory manuals.

[0115] A person skilled in the art is familiar with numerous expression systems available for the expression of nucleic acids encoding the proteins of the present invention.

[0116] ​​​​Alternatively, the nucleic acid of the present invention can be expressed in a host cell by (manipulation) turning on the endogenous DNA encoding the antibody of the present invention within the host cell. Such methods are well known in the art as described in U.S. Patent Nos. 5,580,734, 5,641,670, 5,733,746, and 5,733,761, which are hereby incorporated by reference in their entirety. and can be expressed in the host cell. Such methods are described in U.S. Patent Nos. 5,580,734, 5,641,670, 5,733,746, and 5,733,761, and are well known in the art, and these are hereby incorporated by reference in their entirety.

[0117] An example of a cell culture useful for the production of an antibody, a specified portion thereof, or a variant is a mammalian cell. Mammalian cell lines often take the form of a monolayer of cells, but suspensions or bioreactors of mammalian cells can also be used. Many suitable host cell lines capable of expressing intact glycosylated proteins have been developed in the art, including COS-1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-1651), HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e.g., ATCC CRL1610), and BSC-1 (e.g., ATCC CRL-26) cell lines, Cos-7 cells, CHO cells, hep G2 cells, P3X63Ag8.653, SP2 / 0-Ag14, 293 cells, HeLa cells, etc., which are readily available, for example, from the American Type Culture Collection, Manassas, Va (www.atcc.org). Suitable host cells include cells of lymphoid origin such as myelomas and lymphocytes. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC accession number CRL-1580) and SP2 / 0-A 1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-16 51), HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e.g., ATCC CRL1610), and BSC-1 (e.g., ATCC CRL-26) cell lines, Cos-7 cells, CHO cells, hep G2 cells, P3X63Ag8.653, SP2 / 0-Ag14, 293 cells, HeLa cells, etc., which are readily available, for example, from the American Type Culture Collection, Manassas, Va (www.atcc.org). Suitable host cells include cells of lymphoid origin such as myelomas and lymphocytes. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC accession number CRL-1580) and SP2 / 0-A / 0-Ag14, 293 cells, HeLa cells, etc., which are readily available, for example, from the American Type Culture Collection, Manassas, Va (www.atcc.org). Suitable host cells include cells of lymphoid origin such as myelomas and lymphocytes. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC accession number CRL-1580) and SP2 / 0-A (www.atcc.org). Suitable host cells include cells of lymphoid origin such as myelomas and lymphocytes. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC accession number CRL-1580) and SP2 / 0-A It is g14 cells (ATCC accession number CRL-1851). In a particularly preferred embodiment, the recombinant cell is P3X63Ab8.653 or SP2 / 0-Ag14 cells.

[0118] The expression vectors of these cells include an origin of replication, a promoter (e.g., late or early SV 40 promoter, CMV promoter (U.S. Patent Nos. 5,168,062 and 5,3 85,839), HSV tk promoter, pgk (phosphoglycerate kinase) promoter, EF-1α promoter (U.S. Patent No. 5,266,491), at least one human immunoglobulin promoter, enhancer, and / or ribosome binding site position, RNA splice site, polyadenylation site (e.g., SV40 large T Ag poly addition site), and processing information sites such as transcription termination sequences, etc., but are not limited thereto, and may include one or more of the expression control sequences. For example, see Ausubel et al. above and Sambrook et al. above. Other cells useful for the production of the nucleic acids or proteins of the present invention are known and / or are available, for example, from the American Type Cu lture Collection Catalogue of Cell Lines and Hybridomas (www.atcc.org) or other known or commercially available sources.

[0119] When eukaryotic host cells are utilized, typically a polyadenylation or transcription termination sequence is incorporated into the vector. An example of a termination sequence is the polyadenylation sequence from the bovine growth hormone gene. Sequences for accurate splicing of transcription can also be included as well. An example of a splicing array is the VP1 intron from SV40 (Sprague , et al., J. Virol. 45:773-781 (1983)). In addition, as is known in the art, gene sequences for controlling replication in a host cell can be incorporated into a vector.

[0120] Purification of antibodies. Anti-TNF antibodies can be recovered and purified from recombinant cell cultures by well-known methods such as protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography, but are not limited thereto. High performance liquid chromatography ("HPLC") can also be used for purification. For example, see, e.g., Chapters 1, 4, 6, 8, 9, 10 of Colligan, Current Protocols in Immunology or Current Protocols in Protein Science, John Wiley & Sons, NY, NY (1997-2001), each of which is hereby incorporated by reference in its entirety.

[0121] The antibodies of the present invention include naturally purified products, products of chemical synthesis procedures, and products produced recombinantly from eukaryotic hosts including, for example, yeast, higher plants, insects, and mammalian cells. Depending on the host used in the recombinant production procedure, the antibodies of the present invention may be glycosylated. ​​​​​​​​​​​​​​It may or may not be glycosylated even after glycosylation, but glycosylation is preferred. Such methods are described in the above Sambrook, Sections 17.37-17.42, the above Aus ubel, Chapters 10, 12, 13, 16, 18, and 20, the above Colligan, P rotein Science, Chapters 12-14, and many other standard laboratory manuals and are hereby incorporated by reference in their entirety.

[0122] Anti-TNF antibody All of the heavy chain variable CDR regions of SEQ ID NOs: 1, 2, and 3 and / or SEQ ID NOs: 4, 5, and 6 The isolated antibodies of the present invention containing all of the light chain variable CDR regions are encoded by any suitable polynucleotide, the amino acid sequences of the antibodies disclosed herein, or any isolated or prepared antibodies. Preferably, the human antibody or antigen-binding fragment binds to human TNF, thereby partially or substantially neutralizing at least one biological activity of the protein. At least one biological activity of at least one TNF protein or fragment can be partially or preferably substantially neutralized by an antibody or a specified portion or variant thereof that binds to the protein or fragment, thereby inhibiting the activity mediated through the binding of TNF to the TNF receptor, or through other TNF-dependent or intervening mechanisms. As used herein, the term "neutralizing antibody" refers to an antibody that can inhibit TNF-dependent activity by about 20-12 0%, preferably at least about 10, 20, 30, 40, 50, 55, 60, 65, 70 %, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100% or more, depending on the assay. TNF-dep endent activity. The ability of an anti-TNF antibody to inhibit survival activity is preferably described herein and / or evaluated by at least one suitable TNF protein or receptor assay known in the art. The human antibodies of the invention may be of any class (IgG, IgA, IgM, IgE, IgD, etc.) or isotype and may contain a κ or λ light chain. In one embodiment, the human antibody contains an IgG heavy chain or defined fragment, e.g., at least one isotype of IgG 1, IgG2, IgG3 or IgG4. Antibodies of this type are described herein and / or known in the art, at least one human light chain (e.g., IgG, IgA) and IgM (e.g., γ1, γ2, γ3 γ4) transgenes, and can be prepared by using transgenic mice or other transgenic non-human mammals. In another embodiment, the anti-human TNF human antibody contains an IgG1 heavy chain and an IgG1 light chain.

[0123] As used herein, the term "antibody" or "antibodies" includes biosimilar antibody molecules approved under the Biologics Price Competition and Innovation Act of 2009 (BPCI Act) and similar worldwide laws and regulations. Under the BPCI Act, an antibody is "very similar" to a reference product despite minor differences in clinically inactive components, and data indicating that "it is expected" to obtain clinical results equivalent to the reference product in terms of safety, purity, and potency may demonstrate that it is biosimilar (Endocrine Practice: February 2018, Vol. 24, No. 2, pp. 195- ​​​​​(204). These biosimilar antibody molecules provide a shortened approval pathway, whereby the applicant relies on the clinical data of the innovator's reference product to secure legal approval. Based on the success of clinical trials, compared to the original innovator reference antibody approved by the FDA, the biosimilar antibody molecules are referred to herein as "biological follow-on products". As presented herein, SIMPONI® (golimumab) is the original innovator reference anti-TNF antibody approved by the FDA based on the success of clinical trials. Golimumab has been on the US market since 2009.

[0124] Exemplary sequences Exemplary anti-TNFα antibody sequences, e.g., SIMPONI® (golimumab) Each heavy chain CDR (HCDR) and light chain CDR (LCDR) is underlined in the heavy and light chains of golimumab (defined by Kabat).

[0125]

Chemical formula

[0126]

Chemical formula

[0127] At least one antibody of the present invention binds to at least one specific epitope specific for at least one TNF protein, subunit, fragment, moiety, or any combination thereof. This at least one epitope can comprise at least one antibody-binding region comprising at least a portion of this protein, and this epitope is preferably , at least one extracellular portion, soluble portion, hydrophilic portion, external portion, or cytoplasmic granule portion of this protein. At least one identified epitope is at least 1 to 3 amino acids relative to the entire identified portion of the adjacent amino acids of SEQ ID NO: 9 can include any combination of at least one amino acid sequence.

[0128] Generally, the human antibodies or antigen-binding fragments of the present invention have at least one human complementarity-determining region (CDR1, CDR2, and CDR3) or a variant of at least one heavy chain variable region, and at least one human complementarity-determining region (CDR1, CDR2, and CDR3) or at least one variant of the light chain variable region, and an antigen-binding region. As a non-limiting example, an antibody or the antigen-binding portion or variant can include at least one of the heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 3 and / or the light chain CDR3 having the amino acid sequence of SEQ ID NO: 6. In certain embodiments, the antibody or antigen-binding fragment has an antigen-binding region that includes at least a portion of the corresponding CDR1, 2, and / or 3 amino acid sequences (e.g., SEQ ID NOs: 1, 2, and / or 3) of at least one heavy chain CDR (i.e., CDR1, CDR2, and / or CDR3). In another specific embodiment, the antibody or antigen-binding portion or variant can have an antigen-binding region that includes at least a portion of the corresponding CDR1, 2, and / or 3 amino acid sequences (e.g., SEQ ID NOs: 4, 5, and / or 6) of at least one light chain CDR (i.e., CDR1, CDR2 and / or CDR3). In a preferred embodiment, the three heavy chain CDRs and three light chain CDRs of the antibody or antigen-binding fragment is at least one corresponding CDR of mAb TNV148, TNV14, TNV15, TNV1 96, TNV118, TNV32, and TNV86 described herein. Such antibodies can be prepared and expressed using conventional techniques related to recombinant DNA technology by preparing and expressing a nucleic acid molecule (i.e., one or more) encoding the antibody, or can be prepared by chemically bonding various parts of the antibody (e.g., CDR, framework) together using any other suitable method by using conventional techniques. The anti-TNF antibody can comprise at least one of a heavy or light chain variable region having a defined amino acid sequence. For example, in a preferred embodiment, the anti-TNF antibody optionally comprises at least one of a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and / or at least one light chain variable region having the amino acid sequence of SEQ ID NO: 8. An antibody that binds to human TNF and comprises a defined heavy or light chain variable region can be prepared using suitable methods, such as, for example, phage display (Katsube, Y., et al., Int J Mol. Med, 1( 5):863-868(1998)) or methods employing transgenic animals, which are known in the art and / or described herein. For example, a transgenic mouse comprising a functionally rearranged human immunoglobulin heavy chain transgene and a transgene comprising DNA from a human immunoglobulin light chain locus capable of undergoing functional rearrangement can be immunized with human

[0129] TNF or a fragment thereof to induce the production of antibodies. If desired, the antibody ​​​​​​​​​​​The producing cells can be isolated and, as described herein and / or as known in the art, hybridomas or other immortalized antibody-producing cells can be prepared. Alternatively, an antibody, a specified portion or a variant can be expressed in a suitable host cell using the coding nucleic acid or a portion thereof. The present invention also relates to antibodies, antigen-binding fragments, immunoglobulin chains and CDRs containing amino acids in sequences that are substantially the same as the amino acid sequences described herein. Preferably, such antibodies or antigen-binding fragments and antibodies containing such chains or CDRs have high affinity (e.g., K is about 10 M or less) and can bind to human TNF. Amino acid sequences that are substantially the same as the sequences described herein include sequences containing conservative amino acid substitutions and amino acid deletions and / or insertions. Conservative amino acid substitutions refer to replacing a first amino acid with a second amino acid having chemical and / or physical properties similar to those of the first amino acid (e.g., charge, structure, polarity, hydrophobicity / hydrophilicity). Conservative substitutions include replacing one amino acid with another amino acid within the following groups: lysine (K), arginine (R) and histidine (H); aspartate (D) and

[0130] glutamate (E); asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; alanine (A), valine (V ), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine (C), and glycine (G); F and can bind to human TNF. As amino acid sequences that are substantially the same as the sequences described herein, sequences containing conservative amino acid substitutions and amino acid deletions and / or insertions are exemplified. Conservative amino acid substitutions D is about 10 -9 M or less) and can bind to human TNF. As amino acid sequences that are substantially the same as the sequences described herein, sequences containing conservative amino acid substitutions and amino acid deletions and / or insertions are exemplified. Conservative amino acid substitutions include replacing a first amino acid with a second amino acid having chemical and / or physical properties similar to those of the first amino acid (e.g., charge, structure, polarity, hydrophobicity / hydrophilicity). Conservative substitutions include replacing one amino acid with another amino acid within the following groups: lysine (K), arginine (R) and histidine (H); aspartate (D) and glutamate (E); asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; alanine (A), valine (V ), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine (C), and glycine (G); F include replacing one amino acid with another amino acid within the following groups: lysine (K), arginine (R) and histidine (H); aspartate (D) and glutamate (E); asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; alanine (A), valine (V ), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine (C), and glycine (G); F ), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine (C), and glycine (G); F , W, and Y; C, S, and T.

[0131] Amino acid codes. The amino acids constituting the anti-TNF antibody of the present invention are often abbreviated. Amino acid notations can be indicated by its one-letter code, its three-letter code, name, or the codon(s) of three nucleotides by which the amino acid is represented, which is well understood in the art (see Alberts, B., et al., Molecular Biology of The Cell, Third Ed., Garland Publishing, Inc., New York, 1994).

[0132] [Table 1]

[0133] The anti-TNF antibody of the present invention may include substitutions, deletions, or additions of one or more amino acids by either natural mutations or human operations as specified herein. Of course, the number of amino acid substitutions that can be made by those skilled in the art depends on a number of factors including those described above. Generally speaking, for any given anti-TNF antibody, fragment, or variant, the number of amino acid substitutions, insertions, or deletions, as specified herein, is 40, 30, 2 0, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6,

[0134] 5, 4, 3, 2, 1, for example, 1 - 30, or any range or value within this that does not exceed this. The amino acids within the anti-TNF antibody of the present invention that are functionally essential can be subjected to site-directed mutagenesis or 5, 4, 3, 2, 1, for example, 1 - 30, or any range or value within this that does not exceed .

[0135] ​can be determined by methods known in the art, such as alanine scanning mutagenesis (e.g., Ausubel, supra, Chapters 8, 15; Cunningham and Wells, Science 244:1081-1085 (1989) ). In the latter procedure, mutations with single alanine are introduced for each residue within the molecule. Next , the resulting mutant molecules are tested for biological activities, such as, but not limited to, at least one TNF neutralizing activity. Sites that are crucial for antibody binding can also be identified by structural analysis such as crystallization, nuclear magnetic resonance or photoaffinity labeling (Smith, et al., J. Mol. Biol. 224:8 99-904 (1992) and de Vos, et al., Science 255 :306-312 (1992)). The anti-TNF antibodies of the present invention can include, but are not limited to, at least one moiety, sequence or combination selected from one to all of at least one of the adjacent amino acids of SEQ ID NOs: 1, 2, 3, 4, 5, 6 . The anti-TNF antibodies can further optionally include at least one polypeptide of 70-100% of at least one of the adjacent amino acids of SEQ ID NOs: 7, 8 .

[0136]

[0137]

[0138] In one embodiment, the amino acid sequence of the immunoglobulin chain or a portion thereof (e.g., variable region, CDR) has about 70-100% identity (e.g., 70, 71, 72, 73, 74, 75, 76 with the amino acid sequence of the corresponding chain of at least one of SEQ ID NOs: 7, 8 and ). 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 9 0, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or any range or value therein). For example, the amino acid sequence of the light chain variable region can be compared with the sequence of SEQ ID NO: 8, or the amino acid sequence of the heavy chain CDR3 can be compared with SEQ ID NO: 7. Preferably, 70-100% amino acid identity (i.e., 90, 91, 9 2, 93, 94, 95, 96, 97, 98, 99, 100, or any range or value therein) is determined using suitable computer algorithms as known in the art.

[0139] Exemplary heavy and light chain variable region sequences are shown in SEQ ID NOs: 7 and 8. The antibody of the present invention or its identified variants can include any number of adjacent amino acid residues from the antibody of the present invention, and the number is selected from the group of integers consisting of 10-100% of the number of adjacent residues in the anti-TNF antibody. Optionally, this subsequence of adjacent amino acids is at least about 10, 20, 30 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 1 50, 160, 170, 180, 190, 200, 210, 220, 230, 240, 2 50, or more amino acids in length, or any range or value therein. Further, the number of such subsequences can be any integer selected from the group consisting of 1-20, such as at least 2, 3, 4, or 5. As will be apparent to those skilled in the art, the present invention includes at least one biologically active antibody of the present invention. Biologically active antibodies are natural (non-synthetic), endogenous or related and

[0140] As will be apparent to those skilled in the art, the present invention includes at least one biologically active antibody of the present invention. Biologically active antibodies are natural (non-synthetic), endogenous or related and synthetic (non-natural), exogenous or unrelated antibodies having the same or substantially the same biological activity as the antibodies of the present invention. ​​​​At least 20%, 30% or 40% of that of the known antibody, and preferably at least 50%, 60% or 70%, and most preferably at least 80%, 90% or 95% The enzyme has a specific activity of 1000%. The assay and quantitative measurement of enzyme activity and substrate specificity are , are well known to those skilled in the art.

[0141] In another aspect, the present invention provides a method for the preparation of a compound according to any one of the preceding claims, wherein the compound is modified by the covalent attachment of an organic moiety. The present invention relates to human antibodies and antigen-binding fragments that have been modified to have improved pharmacokinetic properties. (e.g., increased serum half-life in vivo) to produce antibodies or antigen-binding fragments. The organic moiety can be a linear or branched hydrophilic polymer group, a fatty acid group, or a fatty acid ester. In certain embodiments, the hydrophilic polymeric group can be a tertiary ether group. 0 to about 120,000 daltons, and polyalkane glycols (e.g., polyethylene glycols). Polyethylene glycol (PEG), polypropylene glycol (PPG), carbohydrate polymers, It may be an amino acid polymer or polyvinylpyrrolidone, and may also be a fatty acid group or a fatty acid ester. The alkyl group can contain from about 8 to about 40 carbon atoms.

[0142] The modified antibodies and antigen-binding fragments of the invention are directly or indirectly covalently attached to the antibody. The antibody or antigen-binding fragment of the present invention may contain one or more organic moieties that are attached to the antibody or antigen-binding fragment of the present invention. Each organic moiety is independently a hydrophilic polymeric group, a fatty acid group, or a fatty acid ester group. As used herein, the term "fatty acid" refers to mono- and di-carboxylic acids. As used herein, the term "hydrophilic polymeric group" includes octane, It refers to an organic polymer that is more soluble in water than in water. For example, polylysine is It has a higher solubility in water than [the comparison object]. Therefore, an antibody modified by a covalent bond of polylysine is included in the present invention. Suitable hydrophilic polymers for modifying the antibody of the present invention can be linear or branched, for example, polyalkane glycols (e.g., PEG, monomethoxy-poly ethylene glycol (mPEG), PPG, etc.), carbohydrates (e.g., dextran, ce llulose, oligosaccharides, polysaccharides, etc.), polymers of hydrophilic amino acids (e.g., polylysine, po lyarginine, polyaspartic acid, etc.), polyalkane oxides (e.g., polyethylene oxide, polypropylene oxide, etc.), and polyvinyl pyrrolidone. Preferably, the hydrophilic polymer for modifying the antibody of the present invention has a molecular weight of about 800 to about 1 50,000 daltons as an individual molecule. For example, PEG 5000 and PEG 20,00 0 can be used, and the subscript is the average molecular weight (daltons) of the polymer. The hydrophilic polymer group can be substituted with 1 to about 6 alkyl groups, fatty acid groups or fatty acid ester groups. Hydrophilic polymers substituted with fatty acid or fatty acid ester groups can be prepared by using suitable methods. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer. Hydrophilic polymers substituted with fatty acid or fatty acid ester groups can be prepared by using suitable methods. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer.

[0143] Saturated or may contain one or more unsaturated units. Suitable fatty acids and fatty acid esters for modifying the antibody of the present invention Saturated or may contain one or more unsaturated units. Suitable fatty acids and fatty acid esters for modifying the antibody of the present invention Examples of fatty acids include, for example, n-dodecanoate (C 12 , laurate), n-tetradecanoate (C ), myristate), n-octadecanoate (C 14 , stearate), 18 ), n-eicosanoate (C ), arachidate), n-docosanoate (C 20 ), behenate 22 ), n-triacontanoate (C ), n-tetracosanoate (C 30 ), cis-Δ 40 9-octadecenoate (C ), oleate), all cis-Δ5,8,11,14- 18 eicosatetraenoate (C ), arachidonate), octanedioic acid, tetradecane 20 dioic acid, octadecanedioic acid, docosanedioic acid, and the like. Suitable fatty acid esters include monoesters of dicarboxylic acids containing a linear or branched lower alkyl group. The lower alkyl group can contain from 1 to about 12, preferably from 1 to about 6 carbon atoms. Modified human antibodies and antigen-binding fragments can be prepared using suitable methods such as reacting with one or more modifying agents. As used herein, the term "modifying agent"

[0144] means a suitable organic group containing an activating group (e.g., a hydrophilic polymer, a fatty acid, a fatty acid ester). An "activating group" is a chemical moiety or functional group that can react with a second chemical group under appropriate conditions to form a covalent bond between the modifying agent and the second chemical group. For example, amine-reactive activating groups include electrophilic groups such as tosylate, mesylate, halo (chloro, bromo, flu oro, iodo), and N-hydroxysuccinimidyl esters (NHS) . including, etc. As the activating group capable of reacting with thiols, for example, maleimide, iodo cetyl, acryloyl, pyridyldisulfide, 5-thiol-2-nitrobenzoic acid thio ol (TNB-thiol), etc. can be mentioned. The aldehyde functional group can be linked to an amine- or hydra zide-containing molecule, and the azide group can react with a trivalent phosphorus group to form a phospho ramidate or phosphorimide bond. Suitable methods for introducing an activating group into a molecule are known in the art (for example, see Hermanso n, G.T., Bioconjugate Techniques, Academic Press: San Diego, CA (1996)). The activating group can be directly bonded to an organic group (for example, a hydrophilic polymer, a fatty acid, a fatty acid ester), or through a linker part such as a divalent C1-C group (where one or more carbon atoms can be substituted with a heteroatom such as oxygen, nitrogen, or sulfur yellow). Suitable linker 12 parts include, for example, tetraethylene glycol, -(CH2)3-, -NH-(CH2)6 -NH-, -(CH2)2-NH-, and -CH2-O-CH2-CH2-O-CH2-C H2-O-CH-NH-. The modifier containing a linker part can be produced, for example, by forming an amide bond between a free amine and a fatty acid carboxylate by reacting a mono-Boc alkyl diamine (for example, mono-Boc-ethylenediamine, mono-Boc-diamino hexane) with a fatty acid in the presence of 1-ethyl-3- (3-dimethylaminopropyl)carbodiimide (EDC). The Boc protecting group can be removed with trifluoro acetic acid, etc. The modifier containing a linker part can be produced, for example, by forming an amide bond between a free amine and a fatty acid carboxylate by reacting a mono-Boc alkyl diamine (for example, mono-Boc-ethylenediamine, mono-Boc-diamino hexane) with a fatty acid in the presence of 1-ethyl-3- Remove by treatment with acetic acid (TFA) to expose a primary amine that can be linked to another carboxylate as described, or react this with maleic anhydride to cyclize the resulting product to produce an activated maleimide derivative of the fatty acid. (See, for example, International Publication No. 92 / 16221 of Thompson et al., which is hereby incorporated by reference in its entirety.) The modified antibodies of the present invention can be produced by reacting a human antibody or antigen-binding fragment with a modifying agent. For example, the organic moiety can be attached to the antibody in a non-site-specific manner using an amine-reactive modifying agent such as an NHS ester of PEG. It is also possible to prepare a modified human antibody or antigen-binding fragment by reducing the disulfide bonds (e.g., intra-chain disulfide bonds) of the antibody or antigen-binding fragment. At this time, it is possible to react the reduced antibody or antigen-binding fragment with a thiol-reactive modifying agent to produce the modified antibody of the present invention. Modified human antibodies and antigen-binding fragments containing an organic moiety bound to a specific site of the antibody of the present invention are subject to reverse proteolysis (Fisch et al., Bioconjugate Chem., 3

[0145] :147-153 (1992), Werlen et al., Bioconjugat e Chem., 5:411-417 (1994), Kumaran et al., P rotein Sci. 6(10):2233-2241 (1997), Itoh et al., Bioorg.Chem., 24(1):59-68 (1996), Cape :147-153(1992), Werlen et al.,Bioconjugat e Chem.,5:411-417(1994),Kumaran et al.,P rotein Sci.6(10):2233-2241(1997),Itoh et al.,Bioorg.Chem.,24(1):59-68(1996),Cape :147-153(1992),Werlen et al.,Bioconjugat e Chem.,5:411-417(1994),Kumaran et al.,P rotein Sci.6(10):2233-2241(1997),Itoh et al.,Bioorg.Chem.,24(1):59-68(1996),Cape :147-153(1992),Werlen et al.,Bioconjugat Llas et al., Biotechnol. Bioeng., 56(4):456 -463(1997)) and Hermanson, G.T., Bioconjugate Techniques, Academic Press: San Diego, CA( It can be prepared using suitable methods such as those described in 1996).

[0146] Anti-idiotype antibody against the anti-Tnf antibody composition. Monoclonal or chimeric anti-TN In addition to the F antibody, the present invention relates to an anti-idiotype (anti-Id) antibody specific for such an antibody of the present invention. Anti-Id antibodies generally recognize unique determinants related to the antigen-binding region of another antibody. Anti-Id can be prepared by immunizing animals of the same species and genotype as the Id antibody source (e.g., mouse strain) with the antibody or its CDR-containing region. The immunized animal recognizes and responds to the idiotype determinant of the immunizing antibody and produces anti-Id antibodies. Anti-Id antibodies can also be used as an "immunogen" to induce an immune response in yet another animal, generating so-called anti-anti-Id antibodies.

[0147] Anti-Tnf antibody composition. The present invention also provides at least one, at least two, at least three, at least four, at least five, at least six, or more of its anti-TNF antibodies provided in a non-naturally occurring composition, mixture, or form as described herein and / or known in the art. Such a composition is selected from the group consisting of 70-100% of the adjacent amino acids of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, or a specified fragment, domain, or variant thereof. ​​​​At least one or two full-length amino acid sequences of the anti-TNF antibody to be obtained, C and / or N-terminal deletion mutants, domains, fragments, or unnatural-occurring compositions containing specific mutants thereof are included. Preferred anti-TNF antibody compositions are 70-1 00% of the anti-TNF antibodies of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or at least one of their specific fragments, domains, or mutants, containing at least one or two full-length, fragments, domains or mutants as at least one CDR or LBR-containing portion. Even more preferred compositions contain 70-100% of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or at least one of the specific fragments, domains, or mutants thereof, at 40-99%. Such percentages of such compositions are by weight, volume, concentration, molarity, or weight molar concentration as a liquid or dry solution, mixture, suspension, emulsion, or

[0148] colloid, as known in the art or as described herein. The anti-TNF antibody compositions of the present invention further contain at least one anti-TNF antibody for cells, tissues, organs, animals, or patients in need of such regulation, treatment, or therapy, and optionally, at least one TNF antagonist (e.g., TNF antibody or fragment, soluble TNF receptor or fragment, their fusion proteins, or small molecule TNF antagonists, etc., but not limited thereto), antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, etanercept, sodium aurothiomalate, hydroxychloroquine sulfate, leflunomide, Blocking agent, antibacterial agent (e.g., aminoglycoside, antifungal agent, anthelmintic, antiviral agent, carba penem, cephalosporin, fluoroquinolone, macrolide, penicillin, sulfona mide, tetracycline, other antibacterial agents), antipsoriatic agent, corticosteroid, anabolic steroid, diabetes-related drug, mineral, nutritional agent, thyroid drug, vitamin, calcium-related hormone, antidiarrheal agent, antitussive agent, antiemetic agent, antiulcer agent, laxative, anticoagulant, erythropoietin (e.g., epoetin alpha), filgrastim (e.g., G-CSF, Neupogen) , sargramostim (GM-CSF, Leukine), immunizing agent, immunoglobulin, immuno suppressive agent (e.g., basiliximab, cyclosporine, daclizumab), growth hormone, hormone replacement agent, estrogen receptor modulator, mydriatic agent, ciliary muscle regulator, alkylating agent, anti metabolite, mitotic inhibitor, radiopharmaceutical, antidepressant, antimanic drug, antipsychotic drug, anxiolytic drug, hypnotic drug, sympathomimetic agent, stimulant, donepezil, tacrine, asthma drug, beta agonist, inhaled ste roid, leukotriene inhibitor, methylxanthine, cromolyn, epinephrine or similar drug, dornase alpha (Pulmozyme), cytokine or cytokine antagonist selected from at least one of, any suitable and effective amount of a composition or pharmaceutical composition may include at least one of. Non-limiting examples of such cytokines include I L-1 to any of IL-23, but are not limited thereto. Suitable dosages are , well known in the art. For example, Wells et al., eds., P harmacotherapy Handbook, 2 nd Edition, Appl eton and Lange, Stamford, CT(2000), PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing , Loma Linda, CA(2000) is hereby incorporated by reference in its entirety.

[0149] Such anti-cancer agents or anti-infective agents may also include toxin molecules associated with, bound to, co-prescribed with, or used in combination with at least one antibody of the present invention. The toxin can optionally act to selectively kill diseased cells or tissues. Diseased cells can be cancer cells or other cells. Such toxins include, but are not limited to, for example, at least one functional cytotoxic domain selected from at least one of ricin, diphtheria toxin, snake toxin, or bacterial toxin, and can be a purified or recombinant toxin or toxin fragment. The term "toxin" also includes any naturally occurring, mutant or recombinant bacteria or viruses that can cause any condition, including toxic shock that can be fatal in humans and other mammals, including both endotoxins and exotoxins. Such toxins include, but are not limited to, enterotoxigenic Escherichia coli heat-labile enterotoxin (LT), heat-stable enterotoxin (ST), Shigella cell toxin, Aeromonas enterotoxin, toxic shock syndrome toxin-1 ( TSST-1), Staphylococcus enterotoxin A (SEA), B (SEB), or C (S EC), Streptococcus enterotoxin, etc., but are not limited thereto. Such bacteria include enterotoxigenic Escherichia coli (ETEC), enterohemorrhagic Escherichia coli (e.g., serotype 0​​​​​​​​​​ strains of 157:H7), Staphylococcus species (e.g., Staphylococcus aureus, Staphylococcus - pi ogenes), Shigella species (e.g., Shigella dysenteriae, Shigella flexneri, Shigella boydii, and So onnei), Salmonella species (e.g., Salmonella typhi, Salmonella cholerae - suis, Salmo nella enteritidis), Clostridium species (e.g., Clostridium - perfringens, Clostridium - difficile, Clostridium - botulinum), Can pylobacter species (e.g., Campylobacter - jejuni, Campylobacter - fetus ), Helicobacter species (e.g., Helicobacter - pylori), Aeromonas species (e.g ., Aeromonas - sobria, Aeromonas - hydrophila, Aeromonas - caviae) ), Plesiomonas - shigelloides, Yersinia enterocolitica, Vibrio species (e.g., Vibrio cholerae, Vi brio - parahaemolyticus), Klebsiella species, Pseudomonas aeruginosa, and strains of Streptococcus species are included but are not limited thereto. For example, Stein, ed., INTERNAL MEDI CINE, 3rd ed., pp 1 - 13, Little, Brown and Co ., Boston, (1990), Evans et al., eds., Bacter ial Infections of Humans: Epidemiology an d Control, 2d.Ed., pp 239 - 254, Plenum Medic al Book Co., New York(1991), Mandell et al , Principles and Practice of Infectious D iseases, 3d.Ed., Churchill Livingstone, New York(1999). York (1990), Berkow et al, eds., The Merck Manual,16th edition,Merck and Co.,Rahway ,NJ,1992,Wood et al,FEMS Microbiology Immunology,76:121-134(1991), Marrack et a. See, for example, J. Med., Science, 248:705-711 (1990) (these references (the contents of which are incorporated herein by reference in their entirety).

[0150] The anti-TNF antibody compounds, compositions or mixtures of the present invention may further comprise a diluent, binder, stabilizer, buffer, etc. Any additives, such as, but not limited to, buffers, salts, lipophilic solvents, preservatives, adjuvants, etc. The composition may comprise at least one of the following suitable auxiliary agents: Pharmaceutically acceptable auxiliary agents are preferred. Non-limiting examples of and methods for preparing such sterile solutions are well known in the art. For example, see Gennaro, Ed., Remington's Pharmaceut. Physical Sciences, 18 th Edition, Mack Publication Examples of commercially available products include, but are not limited to, 1990. Anti-TNF antibodies, fragments, or antibodies as known in the art or as described herein. A pharma- ceutically acceptable carrier suitable for the mode of administration, solubility, and / or stability of the mutant composition is selected. , can be selected on a daily basis.

[0151] Pharmaceutical additives and excipients useful in the present compositions include, but are not limited to, tannins, Proteins, peptides, amino acids, lipids and carbohydrates (e.g., monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and saccharides containing oligosaccharides, alditols, aldonic acids, derivatized sugars such as esterified sugars, and polysaccharides or sugar polymers), which may be present alone or in combination , and are contained in an amount of 1 to 99.99% by weight or volume, alone or in combination. Exemplary protein additives include serum albumins such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acids / antibody components that can also function in buffering capacity include alanine, glycine, arginine, betaine, histidine , glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine , methionine, phenylalanine, aspartame, and the like. One of the preferred amino acids is glycine.

[0152] Suitable carbohydrate additives for use in the present invention include, for example, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, disaccharides such as lactose, sucrose, trehalose, cellobiose, polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starches, alditols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol),

[0153] myo-inositol, and the like. Preferred carbohydrate additives for use in the present invention are mannitol, trehalose, and raffinose.

[0153] The anti-TNF antibody composition may also contain a buffer or a pH adjuster. Typically, the buffer is a salt prepared from an organic acid or base. Representative buffers include citric acid, ascorbic acid ​Organic acid salts such as salts of acids, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid, and tor Lists, tromethamine hydrochloride, or phosphate buffers. Preferred for use in the present composition The buffer is an organic acid salt such as citrate.

[0154] In addition, the anti-TNF antibody composition of the present invention contains polyvinylpyrrolidone, ficoll (polymer -sugar), dextrate (e.g., 2-hydroxypropyl-β-cyclodextrin, etc. Cyclodextrin), polyethylene glycol, flavoring agent, antibacterial agent, sweetener, antioxidant Agent, antistatic agent, surfactant (e.g., polysorbates such as "TWEEN20" and "TWEEN80" ), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol Terol), and polymer additives / additives such as chelating agents (e.g., EDTA). Obtained.

[0155] These and additional known pharmaceutical additives and / or additives suitable for use in the anti-TNF antibody, partial or variant composition according to the present invention are known in the art, for example "Remington: The Science & Practice of Pha rmacy", 19 ed., Williams & Williams, (1995) th ed., Williams & Williams, (1995) And "Physician's Desk Reference", 52 nd ed, Medical Economics, Montvale, NJ (1998), and these disclosures are hereby incorporated by reference in their entirety into this specification. Preferred carriers or Additive materials are carbohydrates (e.g., monosaccharides and alditols) and buffers (e.g., citric acid ) or polymeric agents.

[0156] As mentioned above, the present invention preferably provides a formulation comprising saline or a selected salt. Stable formulations that are phosphate buffers, as well as preservative-containing storage solutions and formulations, and pharmaceutical US201002326333A1 - Pharmaceutical or veterinary uses comprising at least one anti-TNF antibody in a therapeutically acceptable formulation - Google Patents The present invention provides a multi-use preserved formulation suitable for use in a variety of applications. The preserved formulation comprises at least one pre-existing known, i.e., at least one of phenol, m-cresol, p-cresol, o-cresol, Resole, chlorocresol, benzyl alcohol, phenylmercuric nitrate, phenoxyethanol Alcohol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate) , alkylparaben (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride , benzethonium chloride, sodium dehydroacetate, and thimerosal, or a mixture thereof The composition further comprises a preservative, optionally selected from the group consisting of those known in the art. As in, 0.001 to 5%, or 0.001, 0.003, 0.005, 0.00 9, 0.01, 0.02, 0.03, 0.05, 0.09, 0.1, 0.2, 0.3, 0 .4., 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.3, 4.5, 4.6, Such as, but not limited to, 4.7, 4.8, 4.9, or any range or value therein. Any suitable concentration or mixture may be used, including any range or value therein. This is the case. As non-limiting examples, no preservative is added, 0.1 to 2% m-cresol (e.g., 0. 2, 0.3, 0.4, 0.5, 0.9, 1.0%), about 0.1 to 3% benzyl alcohol (e.g., 0.5, 0.9, 1.1, 1.5, 1.9, 2.0, 2.5%), 0.00 1 to 0.5% thimerosal (e.g., 0.005, 0.01), 0.001 to 2.0% phenol (e.g., 0.05, 0.25, 0.28, 0.5, 0.9, 1.0%), 0.0005 to 1.0% alkyl parabens (plural available) (e.g., 0.00075, 0. 0009, 0.001, 0.002, 0.005, 0.0075, 0.009, 0.01 , 0.02, 0.05, 0.075, 0.09, 0.1, 0.2, 0.3, 0.5, 0. 75, 0.9, 1.0%) and the like can be mentioned.

[0157] As described above, the present invention provides a product comprising a packaging material and at least one vial containing a solution of at least one anti-TNF antibody, optionally with a buffer and / or a preservative formulated in an aqueous diluent, and the packaging material includes a label stating that it can hold such a solution for 1, 2, 3, 4, 5, 6, 9, 12, 18, 20, 24, 30, 36, 40, 48, 54, 60, 66, 72 hours or more. The present invention further includes a product comprising a packaging material, a first vial containing a lyophilized at least one anti-TNF antibody, and a second vial containing an aqueous diluent of a formulated buffer or preservative, and the packaging material includes a label instructing the patient to reconstitute the at least one anti-TNF antibody with the aqueous diluent to form a solution that can be held for 24 hours or more.

[0158] ​​​​​​​At least one anti-TNF antibody used in accordance with the present invention is described herein or may be produced by recombinant means including production from mammalian cells or transgenic preparations known in the art, or may be purified from other biological sources.

[0159] The range of at least one anti-TNF antibody contained in the products of the present invention is an amount that provides a concentration in the range of about 1.0 μg / mL to about 1000 mg / mL when retrieved in the case of a wet / dry system but lower and higher concentrations are also workable, and these concentrations are determined by the intended delivery vehicle, for example, in solution formulations, transdermal patches, lungs, transmucosal, or different from osmotic or micropump methods.

[0160] Preferably, the aqueous diluent optionally further comprises a pharmaceutically acceptable preservative. Preferred preservatives include those selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal or mixtures thereof. The concentration of the preservative used in the formulation is a concentration sufficient to produce an antibacterial effect. Such a concentration varies depending on the selected preservative and can be readily determined by one of ordinary skill in the art.

[0161] Other additives, such as, for example, isotonic agents, buffers, antioxidants, preservative enhancers, may be added to the diluent if desired and preferably. Isotonic agents such as glycerin are commonly used at known concentrations. Preferably, a physiologically tolerated buffer is added to improve the pH ​​​​​​​​Provide control. The formulation is suitable for a wide range of pH ranges, such as from about pH 4 to about pH 10, and preferably from about pH 5 to about pH 9, and most preferably from about 6.0 to about 8.0. It can be done. Preferably, the formulation of the present invention has a pH of about 6.8 to about 7.8. Suitable buffers include phosphate buffers, and most preferably, sodium phosphate, especially phosphate buffered saline (PBS). Other additives, such as Tween 20 (polyoxyethylene (20) sorbitan monolaurate), Tween 40 (polyoxyethylene (20) sorbitan monopalmitate), Tween 80 (polyoxyethylene (20) sorbitan monooleate), Pluronic F68 (polyoxyethylene polyoxypropylene block copolymer), and

[0162] pharmaceutically acceptable solubilizers such as PEG (polyethylene glycol), or nonionic surfactants such as polysorbate 20 or 80 or poloxamer 184 or 188, Pluronic ( registered trademark) polyols, other block copolymers, and chelating agents such as EDTA and EGTA can be optionally added to the formulation or composition to reduce aggregation. These additives are particularly useful when pump or plastic stick containers are used to administer the formulation. The presence of a pharmaceutically acceptable surfactant reduces the tendency of the protein to aggregate. The formulation of the present invention comprises at least one anti-TNF antibody and phenol, m-cresol, p- cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl para cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl para By optionally adding chelating agents such as EDTA and EGTA to the formulation or composition, aggregation can be reduced. These additives are particularly useful when pump or plastic stick containers are used to administer the formulation. The presence of a pharmaceutically acceptable surfactant reduces the tendency of the protein to aggregate. By the presence of a pharmaceutically acceptable surfactant, the tendency of the protein to aggregate is reduced.

[0163] The formulation of the present invention comprises at least one anti-TNF antibody and phenol, m-cresol, p- -cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl para Benzyl (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal or a mixture thereof selected preservatives, and mixing in an aqueous diluent It can be prepared by a process that includes. Mixing at least one anti-TNF antibody and a preservative in an aqueous diluent is a conventional Dissolution and mixing procedures are used. To prepare a suitable formulation, for example, a certain amount of at least one anti-TNF antibody in a buffer solution Is combined with the desired preservative in a buffer solution in an amount sufficient to provide the desired concentration of protein and preservative. Variations Of this process will be recognized by those skilled in the art. For example, the order of addition of components, the use or non-use of additional additives The temperature and pH during formulation preparation are all factors that can be optimized with respect to the administration concentration and means of administration used And are factors that can be optimized. And are all factors that can be optimized with respect to the administration concentration and means of administration used. And are all factors that can be optimized with respect to the administration concentration and means of administration used.

[0164] The claimed formulation can be provided to the patient as a clear solution or as a lyophilized at least one anti-TNF antibody Contained in a second vial containing water, a preservative and / or an excipient, preferably A phosphate buffer and / or saline, and a selected salt, and returned to a vial in an aqueous diluent In a dual vial containing the vial of. Either a single solution vial or a dual vial that requires reconstitution can be reused multiple times And can satisfy a single or multiple patient treatment cycles, and thus can provide a more convenient treatment regimen than is currently available And can provide a more convenient treatment regimen than is currently available.

[0165] The claimed product is useful for administration over a period of from immediately to 24 hours or more. Thus Thus, the product claimed by the present invention provides a great benefit to patients. The formulation can optionally be safely stored at a temperature of about 2 to about 40 °C and can maintain the biological activity of the protein for a long period of time, and thus, on the packaging label, it can be indicated that the solution can be held and / or used for 6, 1 2, 18, 24, 36, 48, 72, or 96 hours or more. When using the stored diluent, such label can include use up to 1 to

[0166] 12 months, half a year, one and a half years, and / or two years. A solution of at least one anti-TNF antibody of the present invention can be prepared by a process including mixing at least one antibody in an aqueous diluent. The mixing is performed using conventional dissolution and mixing procedures. To prepare a suitable diluent, for example, a certain amount of at least one antibody in water or a buffer is combined in an amount sufficient to provide the desired concentration of the protein and, optionally, a preservative or buffer. Variations of this process will be recognized by those skilled in the art.

[0167] For example, the order of addition of components, the presence or absence of additional additives, the temperature and pH during formulation preparation are all factors that can be optimized with respect to the administration concentration and means of administration to be used. The claimed product can be provided to patients as a dual vial containing a lyophilized at least one anti-TNF antibody vial that is reconstituted

[0167] either as a clear solution or in a second vial containing an aqueous diluent. Either a single solution vial or a dual vial that requires reconstitution can be reused multiple times for single or multiple patient treatments.can meet the EULAR, and thus provide a more convenient treatment regimen than currently available. Provide.

[0168] The claimed product can be provided indirectly to patients by providing a dual vial containing a vial of lyophilized at least one anti-TNF antibody that is reconstituted with a transparent solution or aqueous diluent in a second vial. The transparent solution in this case may have a volume of up to 1 liter or even more, and a smaller amount of at least one antibody solution can be taken out from this large container one or more times and transferred to a smaller vial, and can be provided to customers and / or patients by a pharmacy or clinic. vial. For pharmacies, clinics, or other such institutions and facilities. The transparent solution in this case may be up to 1 liter or even more in volume, and a smaller amount of at least one antibody solution can be taken out from this large container one or more times and transferred to a smaller vial, and can be provided to customers and / or patients by a pharmacy or clinic. From this large container, a smaller amount of at least one antibody solution can be taken out one or more times and transferred to a smaller vial. And can be provided to customers and / or patients by a pharmacy or clinic. And / or patients.

[0169] Recognized devices including these single vial systems include pen injector devices for delivering the solution, such as B-D (registered trademark) (pen injector device), NOVOPEN (registered trademark) (pen injector device), AUTOPEN (registered trademark) (pen injector device), OPTIPEN (registered trademark) (pen injector device), GENOTROPIN PEN (registered trademark) (pen injector device), HUMATROPEN (registered trademark) (pen injector device), BJECTOR (registered trademark) (pen injector device), Reco-Pen, Humaject, J-tip Needle-Free Injector, Intraj ect, Medi-Ject, for example, Becton Dickensen (Franklin Lakes, NJ, www.bectondickenson.com) Device), NOVOPEN (registered trademark) (pen injector device), AUTOPEN ( Registered trademark) (pen injector device), OPTIPEN (registered trademark) (pen injector Ector device), GENOTROPIN PEN (registered trademark) (pen injector Device), HUMATROPEN (registered trademark) (pen injector device), BI OJECTOR (registered trademark) (pen injector device), Reco-Pen, Hu maject, J-tip Needle-Free Injector, Intraj ect, Medi-Ject, for example, Becton Dickensen (F ranklin Lakes, NJ, www.bectondickenson.com ), Disetronic (Burgdorf, Switzerland, www.di setronic.comBioject,Portland,Oregon(www. bioject.com);National Medical Products, W eston Medical (Peterborough,UK,www.westo n-medical.com), Medi-Ject Corp (Minneapol (www.mediject.com) Approved devices that contain combination vials include HUMATROPEN (registered a cart for delivering the reconstituted solution, such as the POLYPROPIONATE™ (pen-type injector device); A pen-type injector system for reconstituting lyophilized drugs in a ridge is also included. can be.

[0170] The products claimed herein include packaging. The packaging may be required by a regulatory agency. In addition to providing information about the product, the packaging material of the present invention also provides the conditions under which the product can be used. At least one anti-TNF antibody is reconstituted in an aqueous diluent to form a solution, and incubated for a period of 2 to 24 hours or more. Over the course of the period, the instructions stated that this solution should be used for two vials of wet / dry products. For single vial solution products, the label shall state that the solution is The products claimed herein are intended to be used in human medicine for a period of time longer than that required by the present invention. It is useful for product applications.

[0171] The formulations of the present invention comprise at least one anti-TNF antibody and a selected buffer, preferably a biocompatible buffer. by a process which includes mixing saline or a phosphate buffer containing a selected salt. It can be prepared. Mixing in an aqueous diluent of at least one antibody and a buffer is carried out using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a certain amount of at least one antibody in water or a buffer is combined with the desired buffer in an amount of water sufficient to provide the protein and buffer at the desired concentration. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of components, the use or non-use of additional additives, the temperature and pH during formulation preparation are all factors that can be optimized with respect to the administration concentration and means of administration used.

[0172] The claimed stable or preservative formulation can be provided to a patient as a clear solution or as a dual vial containing a lyophilized at least one anti-TNF antibody that is reconstituted in a second vial containing a preservative or buffer and additives. Either single solution vials or dual vials that require reconstitution can be reused multiple times and can satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0173] At least one anti-TNF antibody of any of the stable or preservative formulations or solutions described herein can be administered to a subject by the present invention via various delivery methods such as SC or IM injection, transdermal, transpulmonary , transmucosal, implantation, osmotic pump, cartridge, micropump or other means well known in the art and understood by those skilled in the art.

[0174] Therapeutic applications. The invention also provides a method for modulating or treating at least one TNF-related disease in cells, tissues, organs, animals or patients using at least one dual integrin antibody of the invention, which is known in the art or described herein.

[0175] The invention also provides a method for modulating or treating at least one TNF-related disease in cells, tissues, organs, animals or patients, including but not limited to at least one of obesity, immune-related diseases, cardiovascular diseases, infectious diseases, malignant diseases or neurological diseases.

[0176] The invention also provides rheumatoid arthritis, juvenile, systemic-onset juvenile rheumatoid arthritis, ankylosing spondylitis, ankylosing spondylitis, gastric ulcer, seronegative arthropathy, osteoarthritis, inflammatory bowel disease, ulcerative colitis, systemic lupus erythematosus, antiphospholipid syndrome, iridocyclitis / uveitis / optic neuritis, idiopathic pulmonary fibrosis, systemic vasculitis / Wegener's granulomatosis, sarcoidosis, orchitis / vasectomy repair, allergic / atopic diseases, asthma, allergic rhinitis, dermatitis, allergic contact dermatitis, allergic conjunctivitis, hypersensitivity pneumonia, transplantation, organ transplant rejection, graft-versus-host disease, systemic inflammatory response syndrome, sepsis syndrome, gram-positive bacterial sepsis, gram-negative bacterial sepsis, culture-negative sepsis, fungal sepsis, neutropenic fever, urosepsis, meningococcal sepsis, trauma / bleeding, burns, ionizing radiation exposure, acute pancreatitis, adult respiratory distress syndrome, alcoholic hepatitis, chronic inflammatory lesions, sarcoidosis, Crohn's lesions, sickle cell anemia, diabetes, nephrosis, atopic diseases, allergic reactions, allergic rhinitis, hay fever, perennial rhinitis, conjunctivitis Meningitis, endometriosis, asthma, urticaria, systemic anaphylaxis, dermatitis, pernicious anemia, hemolytic diseases, thrombocytopenia, graft rejection of any organ or tissue, kidney transplant rejection, heart transplant rejection, liver transplant rejection, pancreas transplant rejection, lung transplant rejection, bone marrow transplant (BMT ) rejection, skin allograft rejection, cartilage transplant rejection, bone graft rejection, small intestine transplant rejection, fetal thymus transplant rejection, parathyroid transplant rejection, xenograft rejection of any organ or tissue rejection, allograft rejection, anti-receptor overreaction, Graves' disease, Raynaud's disease, type B insulin resistance diabetes, asthma, myasthenia gravis, antibody-mediated cell damage, type III hypersensitivity reaction, systemic lupus erythematosus, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin symptoms syndrome), polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin symptoms syndrome, antiphospholipid syndrome, pemphigus, scleroderma, mixed connective tissue disease, idiopathic Addison's disease, true diabetes, chronic active hepatitis, primary biliary cirrhosis, vitiligo, vasculitis, post-MI cardiac surgery syndrome, type IV hypersensitivity, contact dermatitis, hypersensitivity pneumonia, allograft rejection, granuloma due to intracellular organisms, drug allergy, metabolic / idiopathic Wilson's disease, hemochromatosis, α-1-antitrypsin deficiency, diabetic retinopathy, Hashimoto's thyroiditis, osteoporosis, primary biliary cirrhosis, thyroiditis, encephalomyelitis, cachexia, cystic fibrosis, neonatal chronic lung disease, chronic obstructive pulmonary disease (COPD), familial hemophagocytic lymphohistiocytosis, dermatological conditions, psoriasis, alopecia, nephrotic syndrome, nephritis, glomerulonephritis, acute renal failure, hemodialysis, uremia, toxicity, preeclampsia, okt3 therapy, anti-cd3 therapy, cytokine therapy, chemotherapy, radiotherapy (for example, weakness, anemia, cachexia, etc. are including, but not limited to, at least one such as chronic salicylate poisoning but not limited to, at least one immune in cells, tissues, organs, animals or patients A method for modulating or treating related diseases is also provided. For example, Merck Manual , 12th - 17th Editions, Merck & Company, Rahway ay, NJ (1972, 1977, 1982, 1987, 1992, 1999), Pharmacotherapy Handbook, Wells et al., eds. Second Edition, Appleton and Lange, Stamford rd, Conn. (1998, 2000) are hereby incorporated by reference in their entirety respectively, each being incorporated by reference in its entirety .

[0177] The present invention relates to cardiac stun syndrome, myocardial infarction, congestive heart failure , stroke, ischemic attack, hemorrhage, arteriosclerosis, atherosclerotic arteriosclerosis, restenosis, diabetic arteriosclerotic disease , hypertension, arterial hypertension, renovascular hypertension, syncope, shock, cardiovascular syphilis , heart failure, cor pulmonale, primary pulmonary hypertension, arrhythmia, atrial ectopic beats, atrial fibrillation, atrial flutter (persistent or paroxysmal) , post - reperfusion syndrome, cardiopulmonary bypass inflammatory response, chaotic or multifocal atrial tachycardia , regular narrow QRS tachycardia, specific arrhythmias , ventricular fibrillation, His bundle arrhythmias, atrioventricular block, bundle branch block , myocardial ischemic disease, coronary artery disease, angina pectoris, myocardial infarction, cardiomyopathy, dilated congestive cardiomyopathy , restrictive cardiomyopathy, valvular heart disease, endocarditis, pericardial disease, cardiac tumors, aortic aneurysm and peripheral arteries Tumors, aortic dissection, aortic inflammation, occlusion of the abdominal aorta and its branches, peripheral vascular disorders, obstructive arterial disorders, peripheral atherosclerotic arterial disease, thromboangiitis obliterans, functional peripheral arterial disorders, Ray naud's phenomenon and disease, acrocyanosis, erythromelalgia, venous disorders, venous thrombosis, varicose veins, arteriovenous fistulas, lymphedema, lipedema, unstable angina, reperfusion injury, post-pump syndrome, ischemia-reperfusion injury, etc., including at least one of but not limited to these, a method for regulating or treating cardiovascular diseases in cells, tissues, organs, animals or patients is also provided. Such a method may optionally include administering an effective amount of a composition or pharmaceutical composition containing at least one anti-TNF antibody to cells, tissues, organs, animals or patients in need of such regulation, treatment or therapy.

[0178] The present invention also relates to acute or chronic bacterial infections, acute and chronic parasitic or infectious processes including bacterial, viral and fungal infections, HIV infection / HIV neuropathy, meningitis, hepatitis (A, B or C whatever), septic arthritis, peritonitis, pneumonia, epiglottitis, Escherichia coli 0157:h7, hemolytic uremic syndrome / thrombotic thrombocytopenic purpura, malaria, dengue hemorrhagic fever, leishmaniasis, Hansen's disease, toxic shock syndrome, streptococcal myositis, gas gangrene, Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, Pneumocystis carinii pneumonia, pelvic inflammatory disease , orchitis / epididymitis, Legionella, Lyme disease, influenza A, Epstein-Barr virus, virus-associated hemophagocytic syndrome, viral encephalitis / aseptic meningitis, etc., including at least one of but not limited to these, in cells, tissues, organs, animals or patients A method for modulating or treating at least one infectious disease is also provided.

[0179] The present invention also relates to leukemia, acute leukemia, acute lymphoblastic leukemia (ALL), B cells, T cells or FAB ALL, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, myelodysplastic syndrome (MDS), lymphoma, Hodgkin's disease, malignant lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, multiple myeloma, Kaposi's sarcoma, colorectal cancer, pancreatic cancer, nasopharyngeal cancer, malignant histiocytosis, malignant paraneoplastic syndrome / hypercalcemia, solid tumors, adenocarcinoma, sarcoma, malignant melanoma, angioma, metastatic disease, cancer-related bone resorption, cancer-related bone pain, etc., including but not limited to at least one of these, and a method for modulating or treating at least one malignant disease in cells, tissues, organs, animals or patients is also provided.

[0180] The present invention also relates to neurodegenerative diseases, multiple sclerosis, migraine, AIDS dementia complex, demyelinating diseases, for example, multiple sclerosis and acute transverse myelitis, extrapyramidal and cerebellar disorders, for example, lesions of the corticospinal system, disorders of the basal ganglia or cerebellum, hyperkinetic movement disorders, for example, Huntington's chorea and senile chorea, drug-induced movement disorders, for example, those induced by drugs that block CNS dopamine receptors, hypokinetic movement disorders, for example, Parkinson's disease, progressive supranuclear palsy, structural lesions of the cerebellum, spinocerebellar degeneration, for example, spinocerebellar ataxia, Friedreich's Gaucher's disease, abetalipoproteinemia, ataxia, telangiectasia, and mitochondrial multisystem disorders), demyelinating core disorders, such as multiple sclerosis, acute transverse myelitis and motor unit disorders, such as neuropathic muscular atrophy (anterior horn cell degeneration, such as amyotrophic lateral sclerosis, infantile spinal muscular atrophy, and juvenile spinal muscular atrophy), Alzheimer's disease, middle-aged Down syndrome, diffuse Lewy body disease, Lewy body type senile dementia disease, Wernicke-Korsakoff syndrome, chronic alcoholism, Creutzfeldt-Jakob disease subacute sclerosing panencephalitis, Hallervorden-Spatz disease, and boxer dementia, etc., including at least one of these but not limited to them, a method for regulating or treating at least one neurological disease in cells, tissues, organs, animals or patients is also provided. Optionally more, an effective amount of a composition or pharmaceutical composition containing at least one TNF antibody or a specified part or variant is administered to cells, tissues, organs, animals or patients in need of such regulation, treatment or therapy. For example, Merck Manua l,16 Edition, Merck & Company, Rahway, NJ(1 992) for reference. th Edition, Merck & Company, Rahway, NJ(1 992) for reference.

[0181] Any method of the present invention may include administering an effective amount of a composition or pharmaceutical composition containing at least one anti-TNF antibody to cells, tissues, organs, animals or patients in need of such regulation, treatment or therapy. Such a method may further include, optionally, co-administration or combination therapy for the treatment of such immune diseases, and this at least one anti-TNF antibody, specified more, an effective amount of a composition or pharmaceutical composition containing at least one TNF antibody or a specified part or variant is administered to cells, tissues, organs, Administration of the part or its variant thereof is at least one TNF antagonist (e.g., TNF antibody or fragment, soluble TNF receptor or fragment, its fusion protein, or small molecule TNF antagonist, etc., but not limited thereto), antirheumatic drug (e.g., methotrexate , auranofin, aurothioglucose, azathioprine, etanercept, sodium aurothiomalate , hydroxychloroquine sulfate, leflunomide, sulfasalazine), muscle relaxant, anesthetic, non-steroidal anti-inflammatory drug (NSAID), analgesic, anesthetic, sedative, local anesthetic, neuromuscular blocker, antibacterial drug (e.g., aminoglycoside, antifungal drug, anthelmintic, anti-viral drug, carbapenem, cephalosporin, fluoroquinolone, macrolide, penicillin , sulfonamide, tetracycline, other antibacterial drugs), anti-psoriatic agent, corticosteroid , anabolic steroid, diabetes-related drug, mineral, nutritional agent, thyroid drug, vitamin , calcium-related hormone, antidiarrheal, antitussive, antiemetic, anti-ulcer agent, laxative, anticoagulant, erythropoietin (e.g., epoetin α), filgrastim (e.g., G-CSF, Neupogen), sargramostim (GM-CSF, Leukine), immunizing agent, immunoglobulin, immunosuppressive drug (e.g., basiliximab, cyclosporine, daclizumab ), growth hormone, hormone replacement agent, estrogen receptor modulator, mydriatic, cycloplegic , alkylating agent, antimetabolite, mitotic inhibitor, radiopharmaceutical, antidepressant, antimanic drug, antipsychotic drug, anxiolytic, hypnotic, sympathomimetic, stimulant, donepezil, tacrolimus, asthma drug, β agonist, inhaled steroid, leukotriene inhibitor, methylxanthine, cromolyn, epinephrine or analog, dornase alfa (Pulmozyme), cytokine or administering, simultaneously, and / or subsequent to administering at least one selected from cytokine antagonists. Suitable dosages are well known in the art. For nd example, see Wells et al., eds., Pharmacotherapy Handbook, 2 nd Edition, Appleton and Lange, Stamford, CT (2000), PDR Pharmacopoeia, Tarasco Pocket Pharmacopoeia 2000, Deluxe Edition,

[0182] TNF antagonists suitable for the compositions, combination therapies, simultaneous administrations, devices and / or methods of the invention (which further comprise at least one antibody, identified portions thereof and variants thereof of the invention) include anti-TNF antibodies, antigen-binding fragments thereof, and receptor molecules that specifically bind to TNF, compounds that block and / or inhibit (iii), and compounds that block and / or inhibit TNF production and / or synthesis, such as MA including, but not limited to, MP kinase inhibitors.

[0183] As used herein, "tumor necrosis factor antibody", "TNF antibody", "TNFα antibody" ", or "fragment", etc., reduce, block, inhibit, suppress or interfere with TNFα activity in vitro, in situ, and / or preferably in vivo. For example, a preferred TNF human antibody of the present invention can bind to TNFα and includes anti-TNF antibodies, antigen-binding fragments thereof, and identified mutants or domains thereof that specifically bind to TNFα. Preferred TNF antibodies or fragments can also reduce, block, suppress, interfere with, prevent and / or inhibit TNF RNA, DNA, or protein synthesis, TNF release, TNF receptor signaling, membrane TNF cleavage, TNF activity, TNF production and / or synthesis. The chimeric antibody cA2 consists of the antigen-binding variable region of a high-affinity neutralizing mouse anti-human TNFα IgG1 antibody called A2 and the constant region of human IgG1 κ immunoglobulin. The human IgG1 Fc region improves the effector function of the antibody of the same species, increases the circulating serum half-life and reduces the immunogenicity of the antibody. The binding activity and epitope specificity of the chimeric antibody cA2 are derived from the variable region of the mouse antibody A2. In certain embodiments, a preferred source of the nucleic acid encoding the variable region of the mouse antibody A2 is the A2 hybridoma cell line.

[0184] The chimeric A2 (cA2) neutralizes the cytotoxic effects of both native and recombinant human TNFα in a dose-dependent manner. From the binding assay of the chimeric antibody cA2 with recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric

[0185] The chimeric A2 (cA2) neutralizes the cytotoxic effects of both native and recombinant human TNFα in a dose-dependent manner. From the binding assay of the chimeric antibody cA2 with recombinant human TNFα, the chimeric antibody cA2 and recombinant human TNFα, the chimeric The affinity constant of the mela antibody cA2 is 1.04 × l0 10 M -1 It was calculated that The preferred method for determining the specificity and affinity of a monoclonal antibody by immunohistochemistry is rlow,et al.,antibodies:A Laboratory Manu al,Cold Spring Harbor Laboratory Press,C old Spring Harbor, New York, 1988, Colligan et al., eds.,Current Protocols in Immuno logy,Greene Publishing Assoc.and Wiley I terscience, New York, (1992-2000), Kozbor et al., Immunol.Today, 4:72-79(1983), Ausub el et al., eds.Current Protocols in Molec ular Biology,Wiley Interscience,New York (1987-2000), and Muller, Meth. Enzymol., 92:58 9-601 (1983), which references are incorporated herein by reference in their entireties. will be incorporated into the subsection.

[0186] In a specific embodiment, the mouse monoclonal antibody A2 is a polypeptide designated c134A. The chimeric antibody cA2 is produced by a cell line called c168A. It is born.

[0187] Further examples of monoclonal anti-TNF antibodies that can be used in the present invention include It has been described in the art (e.g., U.S. Pat. No. 5,231,024, Moll Er, A., et al., Cytokine 2(3):162-169(1990), U.S. Application No. 07 / 943,852 (filed on September 11, 1992), Rathjen e t al., International Publication No. 91 / 02078 (published on February 21, 1991), Rubin et al., EPO Patent Publication No. 0 218 868 (published on April 22, 1987) , Yone et al., EPO Patent Publication No. 0 288 088 (October 26, 1988), Liang, et al., Biochem. Biophys. Res. Co mmunol. 137:847-854(1986), Meager, et al., Hybri doma 6:305-311(1987), Fendly et al., Hybri doma 6:359-369(1987), Bringman, et al., Hyb ridoma 6:489-507(1987) and Hirai, et al., J.I mmunol. Meth. 96:57-62(1987). See these reference texts (which are hereby incorporated by reference in their entirety).

[0188] TNF receptor molecule. Preferred TNF receptor molecules useful in the present invention bind to TNFα with high affinity (e.g., Feldmann et al., International Publication No. 92 / 07076 (published on April 30, 1992), Schall et al., Cell 61:361-370(1 990), and Loetscher et al., Cell 61:351-359( 1990). See these reference texts (which are hereby incorporated by reference in their entirety ), and have low immunogenicity if desired. In particular, 55 kDa (p55 TNF ​The TNF cell surface receptors of 55 kDa (p55 TNF-R) and 75 kDa (p75 TNF-R) are useful in the present invention. Also useful in the present invention are cleaved forms of these receptors that contain the extracellular domain (ECD) of the receptor or a functional portion thereof. (See, for example, Corcoran et al., Eur. J. Biochem . 223:831-840 (1994)). Cleaved forms of TNF receptors containing the ECD have been detected in urine and serum as 30 kDa and 40 kDa TNFα inhibitory binding proteins (Engelmann, H. et al. , J. Biol. Chem. 265:1531-1536 (1990)). TNF receptor multimeric molecules and TNF immunoreceptor fusion molecules, as well as derivatives and fragments or portions thereof, are further examples of TNF receptor molecules useful in the methods and compositions of the present invention. TNF receptor molecules that can be used in the present invention are characterized by the ability to provide good to excellent alleviation of symptoms and low toxicity in treating patients over a long period of time. Low immunogenicity and / or high affinity, as well as other undefined properties, may contribute to the therapeutic results obtained.

[0189] Useful TNF receptor multimeric molecules in the present invention include all or functional portions of the ECDs of two or more TNF receptors linked via one or more polypeptide linkers or other non-peptide linkers, such as polyethylene glycol (PEG). The multimeric molecules may further include a signal peptide of a secreted protein to effect expression of the multimeric molecule. These multimeric molecules and methods for their production are described in U.S. Application No. 08 / 437,533 filed May 9, 1995, the contents of which are incorporated herein by reference in their entirety.

[0190] ​​​​​ TNF immunoreceptor fusion molecules useful in the methods and compositions of the invention include one or more At least one portion of a globulin molecule and all or a functional portion of one or more TNF receptors. These immunoreceptor fusion molecules may be expressed as monomers or hetero- or homo-multimers. Immunoreceptor fusion molecules can also be assembled in a monovalent or multivalent manner. An example of such a TNF immunoreceptor fusion molecule is a TNF receptor / IgG fusion protein. TNF immunoreceptor fusion molecules and methods for their production have been described in the art. (Lesslauer et al., Eur. J. Immunol. 21:2 883-2886(1991), Ashkenazi et al., Proc. Nat. l.Acad.Sci.USA 88:10535-10539(1991), Pepp el et al., J. Exp. Med. 174:1483-1489 (1991), Kolls et al.,Proc.Natl.Acad.Sci.USA 91:2 15-219 (1994), Butler et al., Cytokine 6(6 ):616-623(1994), Baker et al., Eur. J. Immun ol.24:2040-2048(1994), Beutler et al., U.S. Pat. No. 7,851 and U.S. Application No. 08 / 442,133 (filed May 16, 1995), (Each of these references is incorporated herein by reference in its entirety.) Immune Receptor Fusions Methods for producing the molecule are described by Capon et al., U.S. Pat. No. 5,116,964; Capon et al., U.S. Pat. US Patent No. 5,225,538 and Capon et al., Nature 337: It can also be found in 525-531(1989), and these reference documents are incorporated herein by reference in their entirety. into this specification.

[0191] Functional equivalents, derivatives, fragments, or regions of TNF receptor molecules are of a size and sequence sufficient to be functionally similar to the TNF receptor molecules that can be used in the present invention ( for example, binding to TNFα with high affinity and having low immunogenicity), and refer to parts of TNF receptor molecules, or parts of TNF receptor molecule sequences encoding TNF receptor molecules. Functional equivalents of TNF receptor molecules also include modified TNF receptor molecules that are functionally similar to the TNF receptor molecules that can be used in the present invention (for example, binding to TNFα with high affinity and having low immunogenicity). For example, functional equivalents of TNF receptor molecules may contain the "SILE NT" codon, or one or more amino acid substitutions, deletions, or additions (for example, using one acidic amino acid in place of another acidic amino acid, or using one codon encoding the same or different hydrophobic amino acids in place of another codon encoding hydrophobic amino acids). See Ausubel et al., Current Protocol s in Molecular Biology, Greene Publishing Assoc. and Wiley-Interscience, New York (1 987-2000). Assoc. and Wiley-Interscience, New York (1 987-2000) for reference. See also Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley-Interscience, New York (1987-2000).

[0192] Cytokines include any known cytokines. For example, see Copewith Cytokines.com. Cytokine antagonists include any antibody, fragment or mimetic, any soluble receptor, fragment or mimetic, any small molecule antagonist or any combination thereof, including but not limited to these.

[0193] Treatment. Any method of the present invention can include a method for treating a TNF-mediated disorder, which comprises administering a composition or pharmaceutical composition comprising at least one anti-TNF antibody to a cell, tissue, organ, animal or patient in need of such regulation, treatment or therapy. Such a method may further include, if desired, co-administration or combination therapy for the treatment of such immune diseases, and the administration of at least one of such anti-TNF antibodies, the identified portion or variants thereof is at least one TNF antagonist (for example, a TNF antibody or fragment, a soluble TNF receptor or fragment, its fusion protein, or a small molecule TNF antagonist, etc., but not limited to these ), an antirheumatic drug (for example, methotrexate, auranofin, aurothioglucose ), azathioprine, etanercept, sodium aurothiomalate, hydroxychloroquine sulfate ), leflunomide, sulfasalazine), a muscle relaxant, an anesthetic, a non-steroidal anti-inflammatory drug ([[]] NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antibacterial drug (for example, [[[]] an aminoglycoside, an antifungal drug, an anthelmintic drug, an antiviral drug, a carbapenem, a cephalosporin [[[]] ), a fluoroquinolone, a macrolide, a penicillin, a sulfonamide, a tetracycline, [[[]] other antibacterial drugs), an anti-psoriatic agent, a corticosteroid, an anabolic steroid, a diabetes-related [[[]] drug, a mineral, a nutrient, a thyroid drug, a vitamin, a calcium-related hormone, an antidiarrheal drug, an antitussive [[[]] drug, an antiemetic drug, an antiulcer drug, a laxative, an anticoagulant, erythropoietin (for example, epoetin alpha) [[[]] ), filgrastim (for example, G-CSF, Neupogen), sargramostim (G [[[]] ), M-CSF, Leukine), immunochemicals, immunoglobulins, immunosuppressive drugs (e.g., basiliximab, rituximab, cyclosporine, daclizumab), growth hormones, hormone replacement drugs, estrogen receptor regulators, mydriatics, cycloplegics, alkylating agents, antimetabolites, mitotic inhibitors , radiopharmaceuticals, antidepressants, antimanic drugs, antipsychotics, anxiolytics, hypnotics, sympathomimetics, stimulants, donepezil, tacrine, asthma drugs, beta-agonists, inhaled steroids, leukotriene inhibitors, methylxanthines, cromolyn, epinephrine or analogs, dornase alfa (Pulmozyme), cytokines or cytokine antagonists, and further comprising administering at least one selected therefrom before, simultaneously with, and / or after

[0194] As used herein, the term "safe" refers to the ratio of benefit to the acceptable frequency and / or acceptable severity of adverse events (AE) and serious adverse events (SAE) as compared to a comparator such as standard care or another anti-TNF agent for a favorable risk, when it relates to a composition, dose, administration regimen, treatment or method with an anti-TNF antibody of the present invention. Adverse events are unfavorable medical occurrences in patients administered a pharmaceutical. Specifically, safety when relating to a composition, dose, administration regimen, treatment or method with an anti-TNF antibody of the present invention refers to the acceptable frequency and / or

[0195] acceptable severity of adverse events including, for example, infusion reactions, , the terms "effect" and "effective" refer to the effectiveness of a particular composition, dosage, dosing, treatment or method with an anti-TNF antibody of the present invention (e.g., the anti-TNF antibody golimumab). Effectiveness can be measured based on changes during the course of a disease in response to the agent of the present invention. For example, the anti-TNF antibody of the present invention is administered to a patient in an amount and for a time sufficient to cause improvement, preferably sustained improvement, in at least one indicator reflecting the severity of the disorder being treated. To determine whether the amount and time of such treatment is sufficient, various indicators reflecting the degree of the subject's disease, disorder or condition can be evaluated. Such indicators include, for example, clinically recognized indicators of disease severity, symptoms, or manifestation of the disorder of which the subject is afflicted. The degree of improvement is generally determined by a physician or other appropriately trained individual, who may determine based on signs, symptoms, biopsies, or other test results indicating improvement of clinical symptoms, or any other measure of disease activity. For example, the anti-TNF antibody of the present invention can be administered to achieve improvement in the condition of patients associated with ankylosing spondylitis (AS). Improvement in the condition of patients associated with AS can be evaluated using one or more criteria including, for example, the Ankylosing Spondylitis Disease Activity Score (ASDAS), the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath Ankylosing Spondylitis Metrology Index (BASMI), the 36-item Short Form Health Survey Physical Component Summary (SF-36 PCS), the 36-item Short Form Health Survey Mental Component Summary (SF-36 MCS), and / or the results of the Ankylosing Spondylitis Quality of Life (ASQoL) questionnaire. The ASDAS is For example, the anti-TNF antibody of the present invention can be administered to achieve improvement in the condition of patients associated with ankylosing spondylitis (AS). Improvement in the condition of patients associated with AS can be evaluated using one or more criteria including, for example, the Ankylosing Spondylitis Disease Activity Score (ASDAS), the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath Ankylosing Spondylitis Metrology Index (BASMI), the 36-item Short Form Health Survey Physical Component Summary (SF-36 PCS), the 36-item Short Form Health Survey Mental Component Summary (SF-36 MCS), and / or the results of the Ankylosing Spondylitis Quality of Life (ASQoL) questionnaire. The ASDAS is For example, the anti-TNF antibody of the present invention can be administered to achieve improvement in the condition of patients associated with ankylosing spondylitis (AS). Improvement in the condition of patients associated with AS can be evaluated using one or more criteria including, for example, the Ankylosing Spondylitis Disease Activity Score (ASDAS), the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath Ankylosing Spondylitis Metrology Index (BASMI), the 36-item Short Form Health Survey Physical Component Summary (SF-36 PCS), the 36-item Short Form Health Survey Mental Component Summary (SF-36 MCS), and / or the results of the Ankylosing Spondylitis Quality of Life (ASQoL) questionnaire. The ASDAS is The disease activity for use in AS, developed by the Assessment in Ankylosing Spondylitis Society is the disease activity score (DAS). The ASDAS has, for example, an equation with an assessment including total back pain, duration of morning stiffness, peripheral pain / swelling, and patient global assessment. It is calculated using the equation The BASFI is the self-assessment of the subject represented as the average of 10 questions, 8 of which relate to the subject's functional anatomy and 2 of which relate to the subject's ability to cope with daily life The BASMI is an aggregated score calculated by converting the assessment into scores of 5 assessments including lateral lumbar flexion, tragus-wall distance, lumbar flexion, intercondylar distance, and cervical rotation angle. The SF-36 is a questionnaire consisting of 8 multi-item scales that are scored, and the SF-36 PSA and SF-36 MCS are summary scores obtained from the SF-36 that enable comparison of the relative burden of different diseases and the relative benefits of different treatments The ASQoL is a self-administered patient-reported outcome measure consisting of 18 items that require responses to questions related to sleep, mood, motivation, coping ability, daily life activities, autonomy, interpersonal relationships, and the impact of pain on social life When used in this specification, unless otherwise specified, the term "clinically proven" (used independently or to modify the terms "safety" and / or "efficacy") is intended to mean proven by clinical trials that meet the approval criteria of the US Food and Drug Administration, the EMEA, or the corresponding national regulatory agency. For example, a clinical trial demonstrates the effect of a drug When used in this specification, unless otherwise specified, the term "clinically proven" (used independently or to modify the terms "safety" and / or "efficacy") is intended to mean proven by clinical trials that meet the approval criteria of the US Food and Drug Administration, the EMEA, or the corresponding national regulatory agency. For example, a clinical trial demonstrates the effect of a drug When used in this specification, unless otherwise specified, the term "clinically proven" (used independently or to modify the terms "safety" and / or "efficacy") is intended to mean proven by clinical trials that meet the approval criteria of the US Food and Drug Administration, the EMEA, or the corresponding national regulatory agency. For example, a clinical trial demonstrates the effect of a drug

[0196] As used herein, unless otherwise indicated, the term "clinically proven" (used independently or to modify the terms "safety" and / or "efficacy") is used, for example, to modify the terms "clinically safe" and / or "clinically effective". For example, "clinically proven safety" and / or "clinically proven efficacy" means proven by clinical trials that meet the approval criteria of the US Food and Drug Administration, the EMEA, or the corresponding national regulatory agency. For example, a clinical trial demonstrates the effect of a drug As used herein, unless otherwise indicated, the term "clinically proven" (used independently or to modify the terms "safety" and / or "efficacy") means proven by clinical trials that meet the approval criteria of the US Food and Drug Administration, the EMEA, or the corresponding national regulatory agency. For example, a clinical trial Even a randomized double-blind trial of appropriate size used to clinically prove is good.

[0197] Typically, the treatment of the condition is on average a total of 1 per kg of patient body weight per administration in the range of at least about 0.01 to 500 milligrams of at least one anti-TNF antibody, preferably at least about 0.1 to 100 milligrams per kg of patient body weight per single or multiple administrations of an antibody is achieved by administering a safe and effective amount or dosage of at least one anti-TNF antibody composition. Or, an effective serum concentration may include a serum concentration of 0.1 to 5000 μg / mL per single or multiple administrations. Suitable dosages are known to medical practitioners and, of course, depend on the specific disease state, the specific activity of the composition being administered, and the specific patient being treated . In some cases, repeated administration, i.e., providing repeated individual administrations of a specified monitored amount or dose, may be necessary to obtain the desired therapeutic amount, in which case the individual administrations are repeated until the desired daily dose or effect is obtained.

[0198] Preferred dosages are, if desired, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0 .7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 4 0, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53 , 54, 55, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 8 1, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94 , 95, 96, 97, 98, 99 and / or 100 - 500 mg / kg / dose, or any range, value or fraction thereof, or 0. 1, 0.5, 0.9, 1.0, 1.1, 1.2, 1.5, 1.9, 2.0, 2.5, 2. 9, 3.0, 3.5, 3.9, 4.0, 4.5, 4.9, 5.0, 5.5, 5.9, 6. 0, 6.5, 6.9, 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9. 5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 20, 12.5, 12.9, 13.0, 13.5, 13.9, 14.0, 14.5, 15, 15.5, 15 .9, 16, 16.5, 16.9, 17, 17.5, 17.9, 18, 18.5, 18. 9, 19, 19.5, 19.9, 20, 20.5, 20.9, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 7 0, 75, 80, 85, 90, 96, 100, 200, 300, 400, 500, 600 , 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 35 00, 4000, 4500 and / or 5000 μg / mL serum concentration, or any range, value or fraction thereof that may be obtained.

[0199] Alternatively, the dose administered is based on the pharmacodynamic characteristics of the particular agent, as well as its method and route of administration , the age, health status and weight of the recipient, the nature and degree of the symptoms, the type of concurrent treatment, the treatment It may vary depending on known factors such as frequency and desired effects. The dosage of the active ingredient is usually about 0.1 to 100 milligrams per kilogram of body weight. Usually, 1 per administration, 0.1 to 50, preferably 0.1 to 10 milligrams per kilogram, or a sustained-release form is effective for obtaining the desired results.

[0200] As a non-limiting example, the treatment of humans or animals uses single, infusion or repeated administrations, 1 on at least one day out of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 days, or alternatively, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 5 1 or at least one week out of 52 weeks, or alternatively, 1, 2, 3, 4 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or at least one year out of 20 years, or any combination thereof, 0.1 to 100 mg / kg per day such as, for example, 0.5, 0.9, 1.0, 1.1, 1.5, 2 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 1 8, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40 45, 50, 60, 70, 80, 90, or 100 mg / kg of at least one of the present invention It can be provided as a single or periodic dosage of one antibody.

[0201] Dosage forms (compositions) suitable for parenteral administration generally contain from about 0.1 milligram to about 500 milligrams of the active ingredient per unit or container. In these pharmaceutical compositions, the active ingredient is usually present in an amount of about 0.5 to 99.999% by weight based on the total weight of the composition.

[0202] For parenteral administration, the antibody can be formulated as a solution, suspension, emulsion or lyophilized powder, combined with or separately provided with a pharmaceutically acceptable parenteral vehicle. Examples of such vehicles are water, saline, Ringer's solution, dextrose solution, and 1 - 10% human serum albumin. Non - aqueous vehicles such as liposomes and fixed oils can also be used. The vehicle or lyophilized powder can contain additives (e.g., sodium chloride, mannitol for isotonicity; buffers and preservatives for chemical stability) to maintain isotonicity and chemical stability. The formulation is sterilized by known or suitable techniques. Suitable pharmaceutical carriers are described in the latest edition of Remington’s Pharmaceutical Sciences, A. Osol, a standard reference text in this field.

[0203] Alternative administrations. In order to administer a pharmaceutically effective amount of at least one anti - TNF antibody according to the present invention, many known and developed methods of administration can be used according to the present invention. Although pulmonary administration is used in the following description, other modes of administration may be used according to the present invention to obtain suitable results.

[0204] Alternative administrations. Many known and developed methods of administration can be used according to the present invention to administer a pharmaceutically effective amount of at least one anti - TNF antibody according to the present invention. Although pulmonary administration is used in the following description, other modes of administration may be used according to the present invention to obtain suitable results. ​​​​​

[0205] The TNF antibody of the present invention can be used in a carrier as a solution, emulsion, colloid or suspension, or as a dry powder, and can be delivered by inhalation or by any of a variety of devices and methods suitable for administration by other methods described herein or known in the art.

[0206] Parenteral formulations and administration. Formulations for parenteral administration may contain, as common additives, sterile water or physiological saline, polyalkylene glycols such as polyethylene glycol, oils derived from plants, hydrogenated naphthalene, etc. Aqueous or oily suspensions for injection can be prepared by using appropriate emulsifiers or wetting agents and suspending agents according to known methods. Injectables are, for example, non-toxic parenterally administrable diluents such as aqueous solutions, sterile injectable solutions, or suspensions in a solvent. Possible vehicles or solvents that can be used include water, Ringer's solution, isotonic saline, etc., and as a normal solvent or suspending solvent, sterile non-volatile oils can be used. For these purposes, natural or synthetic or semi-synthetic, fatty oils or fatty acids, natural or synthetic or semi-synthetic, monoglycerides or diglycerides or triglycerides, and all kinds of non-volatile oils and fatty acids can be used. Parenteral administration is known in the art, and includes conventional injection means, gas pressurized needleless injection devices such as those described in U.S. Patent No. 5,851,198, and laser perforator devices such as those described in U.S. Patent No. 5,839,446, but are not limited thereto, and these are hereby incorporated by reference in their entirety.

[0207] Alternative delivery. The present invention further relates to the administration of at least one anti-TNF antibody by parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intra-bronchial, intra-abdominal , intra-pouch, intra-cartilage, intra-cavity, intra-cerebellum, intra-ventricular, intra-colon, intra-cervical, intra-gastric, intra-hepatic, intra-myocardial, intra-bone, intra-pelvic, intra-pericardial, intra-abdominal, intra-pleural, intra-prostatic, intra-pulmonary, intra-rectal, intra-renal, intra-retinal, spinal cord, intra-synovial sac, intra-thoracic, intra-uterine, intra-bladder, bolus, intra-vaginal, rectal, intra-oral, sublingual, nasal cavity, or by transdermal means. At least one anti-TNF antibody composition is for parenteral (subcutaneous, intramuscular or intravenous) or any other administration, in particular, for use in the form of a liquid solution or suspension, in particular, in semi-solid forms such as creams and suppositories but not limited to these, for use in vaginal or rectal administration , in forms such as tablets or capsules but not limited to these, for oral or sublingual administration , or in forms such as powders, nasal drops or aerosols, or certain drugs but not limited to these, for intranasal administration, or for either modifying the skin structure or increasing the drug concentration in a transdermal patch using chemical enhancers such as dimethyl sulfoxide (Junginger, et al. In "Drug Permeatio n Enhancement"; Hsieh, D.S., Eds., pp. 59 - 90 ([[]] Marcel Dekker, Inc. New York 1994, which is hereby incorporated by reference in its entirety [[[]] into this specification), or using oxidizing agents that enable the application of formulations containing proteins and peptides to the skin (International Publication No. WO 98 / 53847), or for creating transient transport pathways such as electroporation or applying an electric field to increase the mobility of charged drugs through the skin such as iontophoresis, or sonophoresis such as ultrasonic introduction ​ Using wave application (U.S. Pat. Nos. 4,309,989 and 4,767,402), gels, ointments, lotions, suspensions, patch delivery systems, etc., but not limited thereto, can be prepared transdermally (the above publications and patents are hereby incorporated by reference in their entirety into this specification).

[0208] Pulmonary / intranasal administration. For pulmonary administration, preferably, at least one anti-TNF antibody composition is delivered at an effective particle size to reach the lower airways or sinuses of the lungs. According to the present invention, at least one anti-TNF antibody can be delivered by any of various inhalation or intranasal devices known in the art for administering therapeutic agents by inhalation. The patient's nasal cavity or alveoli can be coated with the aerosolized formulation. These devices include, for example, metered dose inhalers, nebulizers, dry powder generators, atomizers, etc. Other devices suitable for the purpose of transpulmonary or intranasal administration of the antibody are also known in the art. All such devices can use formulations suitable for administration to distribute the antibody in the aerosol. Such aerosols can be composed of either solutions (both aqueous and non-aqueous) or solid particles. Metered dose inhalers such as VENTOLIN® typically use a propellant gas and require actuation during inhalation (see, for example, WO 94 / 16970, WO 98 / 35888). Turbuhaler® (Astra), Rotahaler® (Glaxo), DISKUS® (inhaler) (Glaxo), SPIROS® (inhaler (Glaxo), etc. A metered dose inhaler such as VENTOLIN® typically uses a propellant gas and requires actuation during inhalation (see, for example, WO 94 / 16970, WO 98 / 35888). Turbuhaler® (Astra), Rotahaler® (Glaxo), DISKUS® (inhaler) (Glaxo), SPIROS® (inhaler (Glaxo), etc. ​)(Dura), devices marketed by Inhale Therapeutics s, and dry powder inhalers such as the Spinhaler® powder inhaler (Fisons), etc. End - expiratory - actuated dry powder inhalers use a mixture of powders (U.S. Patent No. 4,668,218 (Astra) ), European Patent No. 237,507 (Astra), International Publication No. 97 / 25086 (Gla xo), International Publication No. 94 / 08552 (Dura), U.S. Patent No. 5,458,135 (I nhale), International Publication No. 94 / 06498 (Fisons), which are hereby incorporated by reference in their entirety into this specification). AERX® (nebulizer) (Aradigm), U LTRAVENT® (nebulizer) (Mallinckrodt), and the Acorn II nebulizer (Marquest Medical Products) (U.S. Patent No. 5,404,871 (Aradigm), International Publication No. 97 / 22376) ( the above - mentioned documents are hereby incorporated by reference in their entirety into this specification) and other nebulizers generate an aerosol from a solution, while metered - dose inhalers, dry - powder inhalers, etc. generate small - particle aerosols. These specific examples of commercially available inhalation devices are intended to represent particular devices suitable for the practice of the present invention and are not intended to limit the scope of the present invention. Preferably, a composition comprising at least one anti - TNF antibody is delivered by a dry - powder inhaler or a nebulizer. The inhalation devices for administering at least one antibody of the present invention have a plurality of desirable features. For example, delivery by an inhalation device is advantageously highly reliable, reproducible, and accurate. The inhalation device can preferably be used to breathe well, for example, with small particles less than about 10 μm, preferably about 1 - 5 μm. (U.S. Patent No. 5,404,871 (Aradigm), International Publication No. 97 / 22376) ( the above - mentioned documents are hereby incorporated by reference in their entirety into this specification) and other nebulizers generate an aerosol from a solution, while metered - dose inhalers, dry - powder inhalers, etc. generate small - particle aerosols. These specific examples of commercially available inhalation devices are intended to represent particular devices suitable for the practice of the present invention and are not intended to limit the scope of the present invention. Preferably, a composition comprising at least one anti - TNF antibody is delivered by a dry - powder inhaler or a nebulizer. The inhalation devices for administering at least one antibody of the present invention have a plurality of desirable features. For example, delivery by an inhalation device is advantageously highly reliable, reproducible, and accurate. The inhalation device can preferably be used to breathe well, for example, with small particles less than about 10 μm, preferably about 1 - 5 μm. These specific examples of commercially available inhalation devices are intended to represent particular devices suitable for the practice of the present invention and are not intended to limit the scope of the present invention. Preferably, a composition comprising at least one anti - TNF antibody is delivered by a dry - powder inhaler or a nebulizer. The inhalation devices for administering at least one antibody of the present invention have a plurality of desirable features. For example, delivery by an inhalation device is advantageously highly reliable, reproducible, and accurate. The inhalation device can preferably be used to breathe well, for example, with small particles less than about 10 μm, preferably about 1 - 5 μm. These specific examples of commercially available inhalation devices are intended to represent particular devices suitable for the practice of the present invention and are not intended to limit the scope of the present invention. Preferably, a composition comprising at least one anti - TNF antibody is delivered by a dry - powder inhaler or a nebulizer. The inhalation devices for administering at least one antibody of the present invention have a plurality of desirable features. For example, delivery by an inhalation device is advantageously highly reliable, reproducible, and accurate. The inhalation device can preferably be used to breathe well, for example, with small particles less than about 10 μm, preferably about 1 - 5 μm. These specific examples of commercially available inhalation devices are intended to represent particular devices suitable for the practice of the present i...

Claims

1. A method for treating active ankylosing spondylitis in a patient, comprising administering an anti-TNF antibody or an antigen thereof to the patient. administering to a patient a composition comprising a binding fragment, said anti-TNF antibody having the sequence A heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:36 and a light chain comprising the amino acid sequence of SEQ ID NO:

37. (LC), wherein the patient is a responder to the treatment and is not treated with a placebo. There was a statistically significant improvement in disease activity by week 8 of treatment compared with patients who received and the improvement is maintained or improved through 52 weeks of treatment, and the disease activity is Mean change from baseline in ankylosing spondylitis quality of life (AS QoL), 36-item short form Mean change from baseline in physical health score (SF-36 PCS), 36-item short-form mental Mean change from baseline in functional health score (SF-36 MCS), mixed-effects repeated measures statistics Mean change from baseline in the model (MOS-SS) and EuroQol-5D visual The mean change from baseline in the EQ-VAS was The method is determined by the response selected.

2. The statistically significant improvement in disease activity by 16 weeks of treatment was measured based on AS QoL. Mean change from line = -5.4 ± 5.0 standard deviation (SD), based on SF-36 PCS Mean change from baseline in SF-36 MCS = 8.5 ± 7.5 SD mean change from baseline in MOS-SS = 6.6 ± 9.1 SD; mean change from baseline in MOS-SS = 6.6 ± 9.1 SD ±7.2 SD, and mean change from baseline in EQ-VAS = 20.3 ± 24.6 S The method of claim 1 , wherein the compound is selected from the group consisting of D.

3. The composition comprises administering the antibody at a dose of 2 mg / kg, and administering the antibody at weeks 0 and 4, and thereafter IV infusion ( IV).

4. Furthermore, methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine 3. The method of claim 2, further comprising administering the composition with or without roquine (HCQ). The method described above.

5. A detectable label or reporter may be added prior to, simultaneously with, or after said administration. , TNF antagonists, antirheumatic drugs, muscle relaxants, narcotics, nonsteroidal anti-inflammatory drugs (NSAIDs) ), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blocking agents, antibacterial agents, antipsoriatic agents, corticosteroids Steroids, anabolic steroids, erythropoietin, immunizing agents, immunoglobulins, Immunosuppressants, growth hormones, hormone replacement drugs, radiopharmaceuticals, antidepressants, antipsychotics, stimulants Stimulants, asthma medications, beta agonists, inhaled steroids, epinephrine or analogues, cytokines or a cytokine antagonist. and administering at least one composition comprising an effective amount of the compound or protein. The method of claim 1.

6. A composition for use in a method for treating active ankylosing spondylitis in a patient, comprising: administering to the patient a composition comprising a TNF antibody or antigen-binding fragment thereof, The anti-TNF antibody has a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 36 and a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:

37. and a light chain (LC) comprising an amino acid sequence thereof, said patient being a responder to said treatment. There was a statistically significant decrease in disease activity by week 8 of treatment compared to placebo-treated patients. and the improvement was maintained or improved through 52 weeks of treatment. The disease activity is measured by a change from baseline in ankylosing spondylitis quality of life (AS QoL). Mean change, mean from baseline in the 36-item Short Form Physical Health Scale (SF-36 PCS) Change, mean change from baseline in the 36-item Short Form Mental Health Scale (SF-36 MCS) , mean change from baseline in mixed-effects repeated measures statistical model (MOS-SS), and E uroQol-5D visual analogue scale (EQ-VAS) score from baseline The composition, wherein the response is determined by a response selected from the group consisting of: mean change

7. The statistically significant improvement in disease activity by 16 weeks of treatment was measured based on AS QoL. Mean change from line = -5.4 ± 5.0 standard deviation (SD), based on SF-36 PCS Mean change from baseline in SF-36 MCS = 8.5 ± 7.5 SD mean change from baseline in MOS-SS = 6.6 ± 9.1 SD; mean change from baseline in MOS-SS = 6.6 ± 9.1 SD ±7.2 SD, and mean change from baseline in EQ-VAS = 20.3 ± 24.6 S The composition of claim 6, wherein the compound is selected from the group consisting of D.

8. The composition comprises a 2 mg / kg dose of the anti-TNF antibody or antigen-binding fragment thereof. The dose was administered over 30 ± 10 minutes at weeks 0 and 4, and then every 8 weeks (q8w).

7. The composition of claim 6, which is administered by intravenous infusion (IV) so as to be administered immediately. Composition.

9. The method comprises administering methotrexate (MTX), sulfasalazine (SSZ) or hydrochloride to a subject. The method further comprises administering the composition with or without cyclohexene (HCQ). A composition for use in the method according to claim 8.

10. The method further comprises the step of adding a detectable label or reporter, TNF antagonist, antirheumatic drug, muscle relaxant, narcotic, nonsteroidal anti-inflammatory drug (NSAIDs), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blocking agents, antibacterials, antipsoriatics agents, corticosteroids, anabolic steroids, erythropoietin, immunizing agents, immunological Globulin, immunosuppressants, growth hormone, hormone replacement drugs, radiopharmaceuticals, antidepressants, Antipsychotics, stimulants, asthma medications, beta agonists, inhaled steroids, epinephrine or analogues, At least one selected from the group consisting of a cytokine and a cytokine antagonist. and administering at least one composition comprising an effective amount of at least one compound or protein.

7. A composition for use in the method of claim 6 further comprising:

11. A method for treating active ankylosing spondylitis in a patient, comprising administering an anti-TNF antibody or an antigen thereof to the patient. the anti-TNF antibody is an antibody of SEQ ID NO:

36. A heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 37 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:

37. wherein said patients are responders to said treatment compared to placebo-treated patients. By week 8 of treatment, there was a statistically significant improvement in disease activity compared to placebo. and the improvement is maintained or improved through 52 weeks of treatment, and the disease activity is Mean change from baseline in quality of life (AS QoL) and 36-item abbreviated physical health scale Mean change from baseline in the SF-36 PCS and the 36-item short-form mental health scale (S Mean change from baseline in F-36 MCS, mixed-effects repeated measures statistical model (MO Mean change from baseline in the EuroQol-5D visual analog scale (SS-SS) and The response is selected from the group consisting of the mean change from baseline in the EQ-VAS scale (EQ-VAS) The method according to claim 1, wherein the answer is determined by the answer.

12. The statistically significant improvement in disease activity by 16 weeks of treatment was measured based on AS QoL. Mean change from line = -5.4 ± 5.0 standard deviation (SD), based on SF-36 PCS Mean change from baseline in SF-36 MCS = 8.5 ± 7.5 SD mean change from baseline in MOS-SS = 6.6 ± 9.1 SD; mean change from baseline in MOS-SS = 6.6 ± 9.1 SD ±7.2 SD, and mean change from baseline in EQ-VAS = 20.3 ± 24.6 S The method of claim 11 , wherein the compound is selected from the group consisting of D.

13. the anti-TNF antibody or antigen-binding fragment thereof is administered at a dose of 2 mg / kg; Administered over 30 ± 10 minutes at weeks 0 and 4, and then every 8 weeks (q8w) 12. The method of claim 11, wherein the drug is administered by intravenous infusion (IV), such that

14. Methotrexate (MTX), sulfasalazine (SSZ) or hydroxychloroquine administering the anti-TNF antibody or antigen-binding fragment thereof with or without (HCQ); The method of claim 13 , further comprising:

15. A detectable label or reporter may be added prior to, simultaneously with, or after said administration. , TNF antagonists, antirheumatic drugs, muscle relaxants, narcotics, nonsteroidal anti-inflammatory drugs (NSAIDs) ), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blocking agents, antibacterial agents, antipsoriatic agents, corticosteroids Steroids, anabolic steroids, erythropoietin, immunizing agents, immunoglobulins, Immunosuppressants, growth hormones, hormone replacement drugs, radiopharmaceuticals, antidepressants, antipsychotics, stimulants Stimulants, asthma medications, beta agonists, inhaled steroids, epinephrine or analogues, cytokines or a cytokine antagonist. and administering at least one composition comprising an effective amount of the compound or protein. The method of claim 11.

16. Anti-TNF antibody or antigen-binding fragment thereof for use in the treatment of active ankylosing spondylitis in a patient The method further comprises administering to the subject a composition comprising an anti-TNF antibody or an antigen-binding fragment thereof. administering to the patient an anti-TNF antibody comprising the amino acid sequence of SEQ ID NO:

36. and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 37, are responders to said treatment, and are more likely to respond to said treatment compared to placebo-treated patients. By week 8 of treatment, a statistically significant improvement in disease activity was observed, which improvement was The disease activity is maintained or improved through 52 weeks of treatment, and the quality of life (A) in ankylosing spondylitis is improved. Mean change from baseline in S QoL, 36-item Short Form Physical Health Scale (SF-36) Mean change from baseline in the PCS, the 36-item Short Form Mental Health Scale (SF-36 MC Mean change from baseline in sigma-sigma-SS (mean ± SD) and mean mean change from baseline in sigma-sigma-SS (mean ± SD) Mean change from baseline and EuroQol-5D visual analogue scale (E The mean change from baseline in the Q-VAS score was determined by a response selected from the group consisting of: The anti-TNF antibody or antigen-binding fragment thereof.

17. The statistically significant improvement in disease activity by 16 weeks of treatment was measured based on AS QoL. Mean change from line = -5.4 ± 5.0 standard deviation (SD), based on SF-36 PCS Mean change from baseline in SF-36 MCS = 8.5 ± 7.5 SD mean change from baseline in MOS-SS = 6.6 ± 9.1 SD; mean change from baseline in MOS-SS = 6.6 ± 9.1 SD ±7.2 SD, and mean change from baseline in EQ-VAS = 20.3 ± 24.6 S The anti-TNF antibody or its antigen-binding flag of claim 16, selected from the group consisting of: Mention.

18. the anti-TNF antibody or antigen-binding fragment thereof is administered at a dose of 2 mg / kg; Administered over 30 ± 10 minutes at weeks 0 and 4, and then every 8 weeks (q8w) 17. The anti-TNF antibody of claim 16, which is administered by intravenous infusion (IV), as in or an antigen-binding fragment thereof.

19. The method comprises administering methotrexate (MTX), sulfasalazine (SSZ) or hydrochloride to a subject. The anti-TNF antibody or its antigen-binding fragment, with or without cyclohexene (HCQ), 20. The method of claim 18, further comprising administering an anti-TNF antibody to or an antigen-binding fragment thereof.

20. The method further comprises the step of adding a detectable label or reporter, TNF antagonist, antirheumatic drug, muscle relaxant, narcotic, nonsteroidal anti-inflammatory drug (NSAIDs), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blocking agents, antibacterials, antipsoriatics agents, corticosteroids, anabolic steroids, erythropoietin, immunizing agents, immunological Globulin, immunosuppressants, growth hormone, hormone replacement drugs, radiopharmaceuticals, antidepressants, Antipsychotics, stimulants, asthma medications, beta agonists, inhaled steroids, epinephrine or analogues, At least one selected from the group consisting of a cytokine and a cytokine antagonist. and administering at least one composition comprising an effective amount of at least one compound or protein. For use in the method according to any one of claims 16 to 19, further comprising an anti-TNF antibody. or an antigen-binding fragment thereof.

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