Method of treating inflammatory bowel disease with combination therapy of antibodies to il-23 and TNF alpha

Abstract

To provide a method of treating inflammatory bowel disorders such as ulcerative colitis.SOLUTION: Provided is a method comprising administering an IL-23 inhibitor such as an anti-IL-23p19 antibody (e.g., guselkumab) and a TNFα inhibitor such as an anti-TNFα antibody (e.g., golimumab).SELECTED DRAWING: None

Description

[Technical Field]

[0001] Reference to an electronically submitted sequence listing This application is filed under the name "JBI6091WOPCT1SEQLIST.TXT" and The sequence listing is in ASCII format and has a size of 18kb, created on May 20, 2020. This includes sequence listings submitted electronically via EFS-Web. The sequence listing provided is a part of the present specification and is incorporated herein by reference in its entirety. do.

[0002] background Crohn's disease (CD) and ulcerative colitis (UC) Inflammatory bowel diseases (IBD), including idiopathic enteritis, epithelial It is characterized by disruption of the immune barrier and microbial intestinal dysbiosis. The use of medications has significantly changed the clinical management of IBD, but many patients have difficulty responding to induction therapy. The short-term remission rate for biologic therapy used as monotherapy without achieving a clinical response is 2. Less than 0%. (2)

[0003] The role of IL-23 in promoting intestinal inflammation was investigated by treatment with a neutralizing anti-IL-23p19 antibody. In mice with a genetic deletion of the p19 subunit of IL-23, This has been demonstrated in several mouse models, demonstrating toxic colitis. 1, 3-5) Genome-wide association studies (GWAS) In the IL-23 receptor gene (IL23R), which is associated with both risk and protection from IBD, (6) In patients with moderate to severe Crohn's disease, two anti-IL-23 polymorphisms were identified. The drugs risankizumab (BI655066) and brazikumab (MEDI2070, A It was recently reported that the results of a phase 2 trial of MG-139 demonstrated efficacy in the treatment of IBD. There may be a role for anti-IL-23 therapy in the treatment of IL-23-associated leukemia, but as observed with anti-TNFα therapy, It is anticipated that some patient populations may not respond completely to IL-23 alone. do.

[0004] Summary of the Invention Treatment of IBD, particularly based on either anti-TNFα antibodies alone or anti-IL-23 antibodies alone There is a need for improved treatment of IBD in patients who do not respond to treatment.

[0005] One aspect of the present invention is a method for treating inflammatory bowel disease in a patient (subject). a first co-therapeutically effective amount of an IL-23 inhibitor and administering a second co-therapeutically effective amount of a TNFα inhibitor. The method is effective for treating inflammatory bowel disease, and the first and second co-therapeutically effective amounts are the same. or different.

[0006] In some embodiments, the inflammatory bowel disease is ulcerative colitis (UC). In embodiments, the inflammatory bowel disease is Crohn's disease. In some embodiments, the subject has previously received an NF-α inhibitor. have been treated with one agent alone, and their inflammatory bowel disease did not go into remission after previous treatment. In embodiments, the subject has previously been treated with an IL-23 inhibitor alone and the inflammatory bowel disease is , which did not go into remission after previous treatment.

[0007] In various embodiments, the IL-23 inhibitor is an anti-IL-23p19 antibody (referred to herein as (also called anti-p19 or anti-IL-23) or an antigen-binding fragment thereof. In various embodiments, the TNFα inhibitor is an anti-TNFα antibody or antigen-binding fragment thereof. In some embodiments, the anti-IL-23p19 antibody is a human antibody or a humanized antibody. In some embodiments, the anti-TNFα antibody is a human antibody or a humanized antibody. Including the body.

[0008] In some embodiments, the IL-23 inhibitor is (i) SEQ ID NO: 1 (CDRH1); heavy chain CDR amino acid sequences of SEQ ID NO: 2 (CDRH2), and SEQ ID NO: 3 (CDRH3), and (ii) SEQ ID NO: 4 (CDRL1), SEQ ID NO: 5 (CDRL2), and SEQ ID NO: 6 ( 100mg / mL of guselkumab antibody (also known as CNTO1959) (Janssen Biot Tech, Inc. under the trademark Tremfya®) or an antigen thereof Binding fragment; 7.9% (w / v) sucrose, 4.0 mM histidine, 6 0.9 mM L-histidine monohydrochloride monohydrate; 0.053% (w / v) polysorbate 80, and the diluent is water at standard conditions.

[0009] Another aspect of the method of the present invention includes administering a pharmaceutical composition, the pharmaceutical composition comprising SEQ ID NO: The amino acid sequence of the guselkumab heavy chain variable region of SEQ ID NO: 7 and the amino acid sequence of the guselkumab light chain variable region of SEQ ID NO: 8 an isolated anti-IL-23 specific antibody having a region-specific amino acid sequence at 100 mg / mL; The composition was 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L- Histidine monohydrochloride monohydrate; containing 0.053% (w / v) polysorbate 80, diluted The solvent is water at normal conditions.

[0010] A further aspect of the method of the present invention includes administering a pharmaceutical composition, the pharmaceutical composition comprising: The heavy chain amino acid sequence of rucumab (SEQ ID NO: 9) and the light chain amino acid sequence of guselkumab (SEQ ID NO: 10) No. 10) 100 mg / mL of an isolated anti-IL-23 specific antibody; pharmaceutical composition 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine thiamine monohydrochloride monohydrate; 0.053% (w / v) polysorbate 80, diluent is the standard state of water.

[0011] The sequence of guselkumab is as follows:

[0012] [Table 1]

[0013] In various embodiments, the TNFα inhibitor comprises the sequence set forth in the SEQ ID NO: , golimumab antibody (Janssen Biotech, Inc. from Simponi (registered trademark) (sold under the trademark) or an antigen-binding fragment thereof.

[0014] Exemplary Anti-TNFα Antibody Sequence—SIMPONI® (Golimumab) CDR determined by Kabat Amino acid sequence of anti-TNFα antibody complementarity determining region heavy chain 1 (CDRH1): (SEQ ID NO: 11 ) SYAMH

[0015] Amino acid sequence of anti-TNFα antibody complementarity determining region heavy chain 2 (CDRH2): (SEQ ID NO: 12 ) FMSYDGSNKKYADSVKG

[0016] Amino acid sequence of anti-TNFα antibody complementarity determining region heavy chain 3 (CDRH3): (SEQ ID NO: 13 ) DRGIAAGGNYYYYGMDV

[0017] Amino acid sequence of anti-TNFα antibody complementarity determining region light chain 1 (CDRL1): (SEQ ID NO: 14 ) RASQSVYSYLA

[0018] Amino acid sequence of anti-TNFα antibody complementarity determining region light chain 2 (CDRL2): (SEQ ID NO: 15 ) DASNRAT

[0019] Amino acid sequence of anti-TNFα antibody complementarity determining region light chain 3 (CDRL3): (SEQ ID NO: 16 ) QQRSNWPPFT Amino acid sequence of the anti-TNFα antibody heavy chain variable region (CDRs are underlined): Number 17)

[0020] [ka]

[0021] Amino acid sequence of the anti-TNFα antibody light chain variable region (CDRs are underlined): Number 18)

[0022] [ka]

[0023] Amino acid sequence of anti-TNFα antibody heavy chain (CDRs are underlined): (SEQ ID NO: 19 )

[0024] [ka]

[0025] Amino acid sequence of the anti-TNFα antibody light chain (CDRs are underlined): (SEQ ID NO: 20 )

[0026] [ka]

[0027] In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody are mixed in a ratio of 1:2 to 2:2. In some embodiments, the anti-TNFα antibody and the anti-IL-1 antibody are administered in a ratio of 0.1:1 (w / w). -23p19 antibody is used in a ratio of 15:1 to 400:1 (w / w), or in a range of 2:1 to 14:1 It is administered within 24 hours.

[0028] In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody are administered at an initial dose of for subsequent doses, administered simultaneously or on the same day, and for subsequent doses, administered 2 or 3 weeks or more apart. In some embodiments, the two antibodies are administered alternately at intervals of 10 min. In some embodiments, the anti-IL-23p antibody and the anti-TNFα antibody are administered sequentially. The 19 antibody and the anti-TNFα antibody are administered within one day of each other.

[0029] In another aspect, a method of reducing colon inflammation in a subject with inflammatory bowel disease is provided. The method comprises administering an anti-inflammatory drug to a subject in an amount effective to reduce the first co-inflammation. administering an L-23p19 antibody and a second, concurrent, anti-TNFR2 antibody in an amount effective to reduce inflammation; and administering an Fα antibody to the subject. This method reduces inflammation in the colon of the subject compared to that of a normal subject. Effective in reducing inflammation to a level comparable to that of the intestine. The amount and the second, concurrent inflammation-reducing effective amount may be the same or different.

[0030] In some embodiments, the inflammation occurs after administration of an anti-IL-23p19 antibody and an anti-TNFα antibody. The disease is minimal or normal in a tissue sample from the colon of the subject. reported that after administration of anti-IL-23p19 and anti-TNFα antibodies, glandular defects were observed in the colon of subjects. In some embodiments, the anti-IL-23p1 After administration of the anti-TNFα antibody and the anti-TNFα antibody, erosions were most frequently observed in tissue samples from the colon of the subjects. In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody After administration of the antibody, mucosal thickness and hyperplasia were minimal or positive in tissue samples from the colon of the subject. In some embodiments, administration of an anti-IL-23p19 antibody and an anti-TNFα antibody Subsequently, the histopathology of the colon is nearly identical (or identical) to that of normal tissue.

[0031] In another aspect, treating inflammatory bowel disease in a subject and reducing weight loss in a subject. The method includes (a) administering a first co-therapeutic and weight loss-reducing effective amount of an anti-inflammatory drug to a subject. (b) administering a second co-treatment and and administering an anti-TNFα antibody or antigen-binding fragment thereof in an amount effective to reduce the severity of the disease, The first co-treatment and weight loss-reducing effective amount and the second co-treatment and weight loss-reducing effective amount are , are the same or different.

[0032] In another aspect, a method for treating inflammatory bowel disease in a human subject is provided, the method comprising: (a) 0.0005 to 0.002 mg / kg of an anti-IL-23p19 antibody or its antigen binding antibody (b) administering 0.020 to 0.125 mg / kg of an anti-TNFα antibody or and administering the antigen-binding fragment thereof.

[0033] In various embodiments, the method is effective in treating inflammatory bowel disease. In some embodiments, the inflammatory bowel disease is ulcerative colitis. The inflammatory bowel disease is Crohn's disease. In some embodiments, the inflammatory bowel disease is unclassified inflammatory bowel disease. In some embodiments, the method is directed to weight loss (e.g., inflammatory bowel disease). It is effective in preventing the weight loss associated with steroid use.

[0034] In another aspect, a method of preventing colon inflammation in a subject with inflammatory bowel disease is provided. The method includes (a) administering a first concurrent inflammation-reducing amount of an IL-23 inhibitor. and (b) administering a second, concurrent inflammation-reducing amount of a TNFα inhibitor. The method includes reducing inflammation in the subject's colon to a level comparable to that of a normal patient's colon. The amount effective to reduce the first simultaneous inflammation and the amount effective to reduce the second simultaneous inflammation are The amounts effective to alleviate the symptoms may be the same or different.

[0035] In one embodiment, guselkumab is administered at an initial intravenous dose of 200 mg, followed by administration at weeks 4 and 8. UC patients at a dose of 200 mg intravenously, followed by a subcutaneous dose of 100 mg every 8 weeks Golimumab was administered subcutaneously at an initial dose of 200 mg and at weeks 2, 6, and 10. Subsequent subcutaneous doses of 0.00 mg were administered to patients with UC. Patients were randomly assigned to receive UC treatment to determine clinical response or remission. Clinical response measured at 12 weeks was assessed by the Mayo score. A reduction from baseline in ayo score of 30% or more and 3 or more points, A reduction in the rectal bleeding subscore (RBS) of ≥1 or an RBS of 0 or 1. Clinical outcome measured at 12 weeks Remission was defined as a Mayo score of ≤2 with no individual subscores >1 Further measures of clinical response are used within the scope of the present invention. [Brief explanation of the drawings]

[0036] [Figure 1A] Figure 1 shows the results of a weight loss analysis performed on mice after treatment with low doses (Figure 1A, 50 μg) of anti-TNFα and anti-IL-23p19 antibodies, alone or in combination. Each line represents the group mean with error bars for standard error (n = 9 antibody-treated, n = 5 PBS control, n = 3 naive control), shown as percent change from day -1 (dotted line). Some error bars are within the symbol size and are not shown. Disease was induced by administration of an anti-CD40 antibody (BioXCell, catalog number BE0016-2, agonist CD40 Ab clone FGK4.55, lot number 5345 / 0515). [Figure 1B]Figure 1B shows the results of a weight loss analysis performed on mice after treatment with high-dose (500 μg) anti-TNFα and anti-IL-23p19 antibodies, alone or in combination. Each line represents the group mean with error bars for standard error (n = 9 antibody-treated, n = 5 PBS control, n = 3 naive control), shown as percent change from day -1 (dotted line). Some error bars are within the symbol size and are not shown. Disease was induced by administration of an anti-CD40 antibody (BioXCell, catalog number BE0016-2, agonist CD40 Ab clone FGK4.55, lot number 5345 / 0515). [Figure 2A] Figure 2B shows the results of histopathological studies performed on the colons of mice treated with low doses (50 μg / mouse) and high doses (500 μg / mouse) of anti-TNFα and / or anti-IL-23p19 antibodies. Disease was induced by administration of anti-CD40 antibodies. [Figure 2B] Figure 2B shows the results of histopathological studies performed on the colons of mice treated with low doses (50 μg / mouse) and high doses (500 μg / mouse) of anti-TNFα and / or anti-IL-23p19 antibodies. Disease was induced by administration of anti-CD40 antibodies. [Figure 3A] Crohn's Evaluation of Response to Ustekinumab Anti-Interleukin-12 / 23 for Induction (CERTIFI) shows the humanized therapeutic signatures of anti-TNFα or anti-IL-23p19 monotherapy from the anti-CD40 model of murine colitis projected onto the human IBD gene expression network. Figure 3A shows the overlap between genes present in the anti-TNFα and anti-IL-23p19 subnetworks, depicted by a Venn diagram. [Figure 3B] Figure 3B shows the largest connected components of the shared anti-TNFα and anti-IL-23p19 subnetworks. [Figure 4A]Figure 4A shows the results of a weight loss analysis performed on female RAG2- / - mice intraperitoneally administered with an isotype control antibody (Figure 4A), an anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 4B), or an anti-TNFα antibody (150 and 15 μg / mouse) (Figure 4C). Disease was induced by administration of an anti-CD40 antibody. Statistical data were generated to compare each group to the isotype control, as shown in Figure 4D. [Figure 4B] Figure 4A shows the results of a weight loss analysis performed on female RAG2- / - mice intraperitoneally administered with an isotype control antibody (Figure 4A), an anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 4B), or an anti-TNFα antibody (150 and 15 μg / mouse) (Figure 4C). Disease was induced by administration of an anti-CD40 antibody. Statistical data were generated to compare each group to the isotype control, as shown in Figure 4D. [Figure 4C] Figure 4A shows the results of a weight loss analysis performed on female RAG2- / - mice intraperitoneally administered with an isotype control antibody (Figure 4A), an anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 4B), or an anti-TNFα antibody (150 and 15 μg / mouse) (Figure 4C). Disease was induced by administration of an anti-CD40 antibody. Statistical data were generated to compare each group to the isotype control, as shown in Figure 4D. [Figure 4D] Figure 4A shows the results of a weight loss analysis performed on female RAG2- / - mice intraperitoneally administered with an isotype control antibody (Figure 4A), an anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 4B), or an anti-TNFα antibody (150 and 15 μg / mouse) (Figure 4C). Disease was induced by administration of an anti-CD40 antibody. Statistical data were generated to compare each group to the isotype control, as shown in Figure 4D. [Figure 5A]Figure 5A shows the results of histopathological studies performed on the colons of female RAG2- / - mice intraperitoneally injected with an isotype control antibody (Figure 5A), anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 5B), or anti-TNFα antibody (150 and 15 μg / mouse) (Figure 5C). Disease was induced by administration of an anti-CD40 antibody. [Figure 5B] Figure 5A shows the results of histopathological studies performed on the colons of female RAG2- / - mice intraperitoneally injected with an isotype control antibody (Figure 5A), anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 5B), or anti-TNFα antibody (150 and 15 μg / mouse) (Figure 5C). Disease was induced by administration of an anti-CD40 antibody. [Figure 5C] Figure 5A shows the results of histopathological studies performed on the colons of female RAG2- / - mice intraperitoneally injected with an isotype control antibody (Figure 5A), anti-IL-23p19 antibody (50, 15, 5, 1.5, 0.5, and 0.15 μg / mouse) (Figure 5B), or anti-TNFα antibody (150 and 15 μg / mouse) (Figure 5C). Disease was induced by administration of an anti-CD40 antibody. [Figure 6A] Figure 6A shows the results of a weight loss analysis performed on mice administered a control antibody (Figure 6A), an anti-TNFα antibody alone (500 μg / mouse) (Figure 6B), an anti-IL-23p19 antibody alone (1.5, 5, or 25 μg / mouse) (Figure 6C), or a combination of an anti-TNFα antibody (500 μg / mouse) and an anti-IL-23p19 antibody (1.5, 5, or 25 μg / mouse) (Figure 6D). Disease was induced by administration of an anti-CD40 antibody. Figure 6E shows a comparison of the data from the different groups. [Figure 6B] Figure 6A shows the results of a weight loss analysis performed on mice administered a control antibody (Figure 6A), an anti-TNFα antibody alone (500 μg / mouse) (Figure 6B), an anti-IL-23p19 antibody alone (1.5, 5, or 25 μg / mouse) (Figure 6C), or a combination of an anti-TNFα antibody (500 μg / mouse) and an anti-IL-23p19 antibody (1.5, 5, or 25 μg / mouse) (Figure 6D). Disease was induced by administration of an anti-CD40 antibody. Figure 6E shows a comparison of the data from the different groups. [Figure 6C]Figure 6A shows the results of a weight loss analysis performed on mice administered a control antibody (Figure 6A), an anti-TNFα antibody alone (500 μg / mouse) (Figure 6B), an anti-IL-23p19 antibody alone (1.5, 5, or 25 μg / mouse) (Figure 6C), or a combination of an anti-TNFα antibody (500 μg / mouse) and an anti-IL-23p19 antibody (1.5, 5, or 25 μg / mouse) (Figure 6D). Disease was induced by administration of an anti-CD40 antibody. Figure 6E shows a comparison of the data from the different groups. [Figure 6D] Figure 6A shows the results of a weight loss analysis performed on mice administered a control antibody (Figure 6A), an anti-TNFα antibody alone (500 μg / mouse) (Figure 6B), an anti-IL-23p19 antibody alone (1.5, 5, or 25 μg / mouse) (Figure 6C), or a combination of an anti-TNFα antibody (500 μg / mouse) and an anti-IL-23p19 antibody (1.5, 5, or 25 μg / mouse) (Figure 6D). Disease was induced by administration of an anti-CD40 antibody. Figure 6E shows a comparison of the data from the different groups. [Figure 6E] Comparison of data from different groups is shown. [Figure 7A] Figure 7 shows the results of histopathological studies performed on the colons of mice administered anti-TNFα antibody alone (500 μg / mouse), murine anti-IL-23p19 antibody alone, or a combination of anti-TNFα antibody (500 μg / mouse) and murine anti-IL-23p19 antibody (anti-IL23p19 antibody concentrations: 1.5 μg (Figure 7A), 5 μg (Figure 7B), or 25 μg (Figure 7C)). Disease was induced by administration of anti-CD40 antibody. [Figure 7B] Figure 7 shows the results of histopathological studies performed on the colons of mice administered anti-TNFα antibody alone (500 μg / mouse), murine anti-IL-23p19 antibody alone, or a combination of anti-TNFα antibody (500 μg / mouse) and murine anti-IL-23p19 antibody (anti-IL23p19 antibody concentrations: 1.5 μg (Figure 7A), 5 μg (Figure 7B), or 25 μg (Figure 7C)). Disease was induced by administration of anti-CD40 antibody. [Figure 7C]Figure 7 shows the results of histopathological studies performed on the colons of mice administered anti-TNFα antibody alone (500 μg / mouse), murine anti-IL-23p19 antibody alone, or a combination of anti-TNFα antibody (500 μg / mouse) and murine anti-IL-23p19 antibody (anti-IL23p19 antibody concentrations: 1.5 μg (Figure 7A), 5 μg (Figure 7B), or 25 μg (Figure 7C)). Disease was induced by administration of anti-CD40 antibody. [Figure 8] Figure 1 shows the results of a network analysis based on humanized colon gene expression signatures of anti-TNFα (500 μg) monotherapy or high-dose anti-IL-23p19 (25 μg) monotherapy crossed with gene expression signatures from the combination therapy (500 μg anti-TNFα and 1.5 μg anti-IL-23p19). This analysis determined whether the molecular response to the combination treatment of anti-TNFα and low-dose anti-IL-23p19 antibody was additive or specific compared to either therapy alone. A unique subnetwork of approximately 200 genes was identified, and the subnetwork was enriched in cell types and pathways involved in fibroblast and extracellular matrix organization, wound repair, and mucosal healing.

[0037] Detailed Description Definition: Unless otherwise specified, technical and scientific terms used in this specification and claims The terms have the same meaning as commonly understood by a person skilled in the art to which this invention pertains. It has one meaning.

[0038] As used in this specification, including the appended claims, the singular forms "a," "an," "an" and "an" are used interchangeably. "d" and "the" refer to their counterparts unless the context clearly indicates otherwise. It contains multiple referents that correspond to the same thing.

[0039] "About" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art. This means that the value is determined by the way it is measured or determined, i.e., the limitations of the measurement system. To some extent, the examples or results may be used in the context of a particular assay, result, or embodiment. Unless expressly stated otherwise elsewhere in the specification, "about" means any material that falls within the scope of the practice of the art. This means that the value is within one standard deviation or 5% of the range, whichever is greater. do.

[0040] When applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, "Administer" and "treat" refer to any treatment or administration to an animal, human, subject, cell, tissue, organ, or biological fluid. "Administration" and "treatment" refer to the contact of an exogenous pharmaceutical, therapeutic, diagnostic, or composition. For example, it can refer to therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of cells involves contact of reagents with the cells as well as reagents with fluids in contact with the cells. "Administration" and "treatment" also include administering or administering to a subject by a reagent, diagnostic, binding composition, or the like. "Treatment" refers to in vitro and ex vivo treatment of cells, e.g., by another cell. "Treatment" means therapeutic treatment, prophylactic or preventative treatment when applied to human, veterinary, or research subjects. "Treatment" refers to the use of a substance in a human subject, a veterinary subject, or a research subject. When applied to a research subject, or to a cell, tissue, or organ, "Treatment of cells" also includes contacting a biological compartment, or physiological fluid, with an agent. For example, in a fluid or colloidal phase, the target, such as the IL-23 receptor, may be contacted. However, it also includes situations where the agonist or antagonist does not contact the cell or receptor. do.

[0041] "Treatment" or "treating" also refers to administering the therapeutic agent to a patient in need thereof using the combinations described herein. It can refer to the oral or topical administration of a therapeutic agent such as a drug. The agent may be used to treat one or more pathological conditions or one or more of the treatments with different therapeutic agents. Adverse reactions are those that reduce the occurrence of such symptoms or side effects to any clinically measurable degree. To prevent or alleviate, whether by preventing, inducing regression, or inhibiting progression Therapeutic agents effective in alleviating any particular pathological condition or side effect are administered in an amount effective to alleviate any particular pathological condition or side effect. The amount of the therapeutic agent (also called a "therapeutically effective amount") will depend on the patient's condition, age, weight, and desired response of the patient. the ability of the therapeutic agent to elicit a response, the patient's overall health, the method, route and dose of administration, and side effects. The duration of treatment may vary depending on factors such as the severity of the condition.

[0042] "Inhibitor," as used herein, refers to any compound that reduces the activity of a target molecule. Specifically, the antagonist of IL-23 or TNFα is, for example, 23 or TNFα by blocking its receptor binding or by other means. By reducing the activity of I (e.g., as measured in a bioassay), It is a drug that reduces the biological activity of L-23 or TNFα.

[0043] As used herein, "anti-IL-23 specific antibody," "anti-IL-23 antibody," "," "antibody portion," or "antibody fragment" and / or "antibody variant," etc., are used to describe antibodies of the present invention. At least one complementarity determining region of the heavy or light chain can be incorporated into CDR) or its ligand-binding portion, heavy chain or Light chain variable region, heavy or light chain constant region, framework region, or any part thereof or at least a portion of the IL-23 receptor or binding protein, Any protein or molecule containing at least a portion of an immunoglobulin molecule, including, but not limited to: Such antibodies may optionally be further linked to specific ligands. For example, but not by way of limitation, such antibodies may be used in vitro, in vivo, and / or in vivo at least one IL-23 activity or binding, or IL-23 activity or binding. -23 receptor activity or binding, modulating, reducing, increasing, antagonizing, stimulating, alleviating, mitigating, blocking, As a non-limiting example, a suitable anti-IL-23 antibody of the present invention The antibody, specified portion, or variant is a polypeptide of at least one IL-23 molecule or a specified portion thereof. Suitable anti-IL-23 antibodies, particularly those capable of binding to IL-23 fragments, variants, or domains thereof, include: The determined portion or variant also optionally possesses at least one IL-23 activity or Such activities or functions can also affect the activity or function of RNA, DNA, or protein synthesis, IL-23 release, IL-23 receptor signaling, membrane IL These include IL-23 cleavage, IL-23 activity, IL-23 production, and / or synthesis. , but not limited to.

[0044] The term "antibody" includes antibody mimetics, or single chain antibodies and fragments thereof. A portion of an antibody or a specific fragment or fragments thereof that mimics the structure and / or function of the antibody. The present invention also includes antibodies, digested fragments thereof, specified portions, and variants thereof, including antibodies or portions thereof. It is further intended that the functional fragments include antigen-binding fragments that bind to mammalian IL-23. For example, antibody fragments capable of binding to IL-23 or a portion thereof include , Fab (e.g., by papain digestion), Fab' (e.g., by pepsin digestion and partial by reduction) and F(ab')2 (e.g., by pepsin digestion), facb (e.g., pFc' (e.g., by pepsin or plasmin digestion), F d (e.g., by pepsin digestion, partial reduction, and reassembly), Fv or scFv (e.g., Examples of fragments include, but are not limited to, fragments obtained by molecular biology techniques.

[0045] Such fragments may be used as known in the art and / or as described herein. Antibodies can be produced by enzymatic cleavage, synthetically, or recombinantly, as described in The present invention relates to an antibody gene in which one or more stop codons have been introduced upstream of the natural stop site. Various truncated forms can also be produced using the fragments. For example, the heavy chain portion of F(ab')2 The combined gene encoding the heavy chain also encodes the CH1 domain and / or hinge region of the heavy chain. The various portions of the antibody can be chemically engineered using conventional techniques to contain DNA sequences encoding the antibody. or can be bound to a continuous protein using genetic engineering techniques. It can be prepared as follows.

[0046] A "humanized antibody" is an antibody in which the antigen-binding site is derived from a non-human species and the variable region framework is derived from a human Humanized antibodies refer to antibodies derived from human immunoglobulin sequences. Since the framework may contain expressed human immunoglobulins or human It need not be an exact copy of the immunoglobulin germline gene sequence.

[0047] A "human antibody" is an antibody in which both the framework and antigen-binding sites are derived from sequences of human origin. The term "antibody" refers to an antibody having a heavy chain variable region and a light chain variable region that are identical to the constant region or part of the constant region. If the constant region comprises a sequence of human origin, the constant region is also derived from a sequence of human origin.

[0048] "Subject" or "patient," when used interchangeably, includes any human or non-human animal. "Non-human animals" include, for example, non-human primates, sheep, dogs, cats, horses, cows, chickens, etc. Includes all vertebrates, including mammals such as birds, amphibians, and reptiles, as well as non-mammals .

[0049] "Tumor necrosis factor," "TNF," or "TNFα" refers to the well-known human tumor necrosis factor-α (TNFα). TNFα is a multifunctional pro-inflammatory cytokine that promotes cartilage and bone degradation. , induction of adhesion molecules, induction of procoagulant activity on vascular endothelial cells, adhesion of neutrophils and lymphocytes Increased blood cholesterol and release of platelet-activating factor from macrophages, neutrophils, and vascular endothelial cells Induce pro-inflammatory pathways that result in tissue damage, such as stimulation of the immune system.

[0050] TNFα is expressed as a soluble protein as well as a type II polypeptide on the surface of cells. Transmembrane TNFα is found in a precursor form called transmembrane TNFα, which is expressed in the TN Residues Ala76 and V are degraded by metalloproteinases such as Fα-converting enzyme (TACE). a177, resulting in the release of a soluble form of TNFα of 157 amino acid residues. Soluble TNFα is a homotrimer of 17 kDa truncated monomers. TNFα also exists as a homotrimer of 26 kDa uncleaved monomers.

[0051] In a first aspect, a method of treating inflammatory bowel disease in a subject is provided. The method comprises: Administer a co-therapeutically effective amount of one IL-23 inhibitor and administering a second co-therapeutically effective amount of a TNFα inhibitor. are effective to treat inflammatory bowel disease, and the first and second co-therapeutically effective amounts are the same. or different.

[0052] The combination of anti-TNFα and anti-IL-23p19 antibodies has been shown to reduce systemic effects as well as intestinal or provide a localized effect on the colon. provides a greater systemic effect than treatment with either the IL-23p19 antibody or anti-IL-23p19 antibody alone This combination has been shown to be an excellent anti-inflammatory agent in the treatment of IBD in humans. Anti-IL-23p19 antibodies can provide therapeutic activity in IBD (e.g., colitis and Although anti-CD40 derivatives can be highly effective in preventing the development of Crohn's disease, While anti-TNFα antibodies cannot block IBD, they offer some protection from IBD. Each antibody and this can provide substantial protection from anti-CD40-induced weight loss. These combinations may have differential effects on local versus systemic inflammation. .

[0053] Various anti-IL-23 antibodies, such as those listed in Table 2, are incorporated herein by reference. U.S. Patent No. 7,491,391 issued February 17, 2009, and U.S. Patent No. 7,491,391 issued April 5, 2018 Any of the anti-IL-23 antibodies described in U.S. Patent Application Publication No. 2018 / 0094052 Either one or the like can be used.

[0054] A variety of anti-TNFα antibodies may be used, for example, No. 7,250,165 issued July 31, 2007, and U.S. Patent No. 7,250,165 issued August 31, 2017, The anti-IL-1 inhibitors described in U.S. Patent Application Publication No. 2017 / 0218092, published on March 3, Any of the 23 antibodies can be used.

[0055] A variety of host animals can be used to produce anti-TNFα antibodies. b / c mice can be used to generate mouse anti-human TNFα antibodies. Antibodies generated in / c mice and other non-human animals are being developed to produce more human-like sequences. They can also be humanized using a variety of techniques to create polypeptides.

[0056] The anti-IL-23 antibody optionally has high affinity binding to IL-23, and optionally The anti-TNFα antibody may optionally be characterized by its ability to inhibit TNFα. and, optionally, low toxicity. , individual components such as variable regions, constant regions, and frameworks, individually and / or Collectively, antibodies, specific antibodies, optionally and preferably, have low immunogenicity. fragments or variants thereof may be used. The high affinity of the hydroxybenzoates, as well as other favorable properties, can contribute to the therapeutic results obtained. "Low immunogenicity" as used herein refers to a level of immunogenicity in less than about 75% of treated patients, or preferably less than about 75% of treated patients. a significant increase in HAHA, HACA, or HAMA response of less than about 50%; and / or , low titers (less than about 300 as measured by double antigen enzyme immunoassay) in treated patients , preferably less than about 100) (Elliott et al. al., Lancet 344:1125-1127 (1994), the entire contents of which are incorporated herein by reference. With respect to anti-IL-23 antibodies, "low immunogenicity" means that the antibody Regarding the recommended course of treatment, in patients treated with anti-IL-23 antibodies, Titratable levels of antibodies to β-lactamase occur in less than 25% of patients treated at the recommended dose , preferably in less than 10% of treated patients. With respect to NFα antibodies, "low immunogenicity" refers to the anti-T Antibodies to anti-TNFα antibodies reach titratable levels in patients treated with anti-TNFα antibodies The frequency of this condition is less than 25% of patients treated at the recommended dose, preferably 10% of patients treated It can also be defined as being less than 10%.

[0057] At least one anti-IL-23 antibody and an anti-TN antibody used in the method according to the present invention Fα can be produced from cell lines, mixed cell lines, immortalized cells, or immortalized cells, as known in the art. It can be produced by a clonal population of, for example, Ausubel, et al. ,ed.,Current Protocols in Molecular Biol. ogy, John Wiley & Sons, Inc., NY, (1987-2001), Sambrook, et al., Molecular Cloning: A Labo ratory Manual,2nd Edition,Cold Spring Ha rbor, NY (1989), Harlow and Lane, Antibodi. es,a Laboratory Manual,Cold Spring Harbo r, NY (1989), Colligan, et al., eds., Curren. t Protocols in Immunology,John Wiley&Son s, Inc., NY (1994-2001), Colligan et al., Cur. rent Protocols in Protein Science,John W. See, e.g., Iley & Sons, NY (1997-2001), each of which is incorporated by reference. The entire contents of which are incorporated herein.

[0058] Anti-IL-23 antibodies and / or anti-TNFα antibodies may also be used in the methods described herein and / or of the present invention. As is known in the art, a repertoire of human antibodies can be produced. Transgenic animals (e.g., mice, rats, hamsters, non-human primates, etc.) Human anti-IL-23 antibody-producing cells may be derived from the human anti-IL-23 antibody-producing cells described herein. They may be isolated from such animals and immortalized using suitable methods, such as those described above.

[0059] The anti-IL-23 antibodies used in the methods described herein also inhibit the production of such antibodies in milk. transgenic animals or mammals such as goats, cows, horses, sheep, and rabbits that and preparing a method for the treatment of IL-23 comprising the steps of: The anti-TNFα antibodies used in the methods described herein can also be used to treat such antibodies. Transgenic animals or mammals such as goats, cows, horses, sheep, rabbits, etc. that produce it in their milk Using nucleic acids encoding at least one anti-TNFα antibody to prepare a mammal Such animals can be prepared using known methods. See, for example, but not limited to, U.S. Patent Nos. 5,827,690 and 5,849,999. No. 92, No. 4,873,316, No. 5,849,992, No. 5,994,61 See, for example, Patent Nos. 6, 5,565,362, and 5,304,489. each of which is incorporated herein by reference in its entirety.

[0060] Anti-IL-23 antibodies have broad affinity (K D ) and can bind to human IL-23 In a preferred embodiment, the human mAb optionally binds to human IL-23 with high affinity. For example, a human mAb can bind to human IL-23 at approximately 10 -7 M and below, For example, but not limited to, 0.1 to 9.9 (or any range or value therein). ) x 10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 or any range or value therein, etc. D can be combined with

[0061] Anti-TNFα antibodies have a wide range of affinities (K D ) and can bind to human TNFα. In a preferred embodiment, the human mAb optionally binds to human TNFα with high affinity. For example, a human mAb can bind to human TNFα at approximately 10 -7 Below M, e.g. For example, but not limited to, 0.1 to 9.9 (or any range or value therein). x10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 also is any range or value within KD can be combined with

[0062] The anti-IL-23 antibody is of the IgG1, IgG2, IgG3, or IgG4 isotype. The anti-TNFα antibody may be of the IgG1, IgG2, IgG3, or IgG4 isotype. It is possible.

[0063] Without wishing to be bound by theory, it is believed that the anti-IL-23p19 antibody and the anti-TNFα antibody The benefits of combining antibodies arise from the changes in gene expression induced by each antibody. As described in Example 1 and at least in Figures 2A and 2B, each antibody At doses that provided similar protection against inflammation (Figure 2, 50 μg anti-IL-23p19 and 5 00 μg anti-TNFα), when comparing IL-23p19 blockade with TNFα blockade, Differential changes in intestinal gene expression were observed in mice. This may also apply to humanized mouse anti-TNFα and anti-IL-23p19 genes. By integrating the signature with human intestinal biopsy gene networks, we identified the genetic makeup of human intestinal tissue. It is possible to focus only on genes that are expressed and altered in response to the antibody. Further context potentially impacting the network is provided by the genes in each signature. Treatment subnetwork containing genes one step removed (i.e., strongly correlated) The individual anti-TNFα and anti-IL-23p19 antibodies can be obtained by generating The network exhibits a unique, single antibody gene signature, and thus is likely to be mediated by both mechanisms. Enabling insight into targeted biology.

[0064] The effectiveness of treatment with the methods described herein can be measured, for example, by measuring the extent of weight loss, nutrient absorption, and the like. , and can be determined by evaluating histopathological studies of tissue samples. Histological studies may reveal one of the following: submucosal edema, inflammation, glandular defects, erosion, mucosal thickening, and hyperplasia. or two or more measurements. Submucosal edema may be measured (e.g., to assess the severity of this change). The thickness from the muscularis mucosae to the inner border of the superficial muscularis (within the non-tangential region that is considered to best represent the Inflammation scoring can be quantified by measuring the amount of macrophage in the colon. This may reflect the degree of phage infiltration, lymphocyte infiltration, and neutrophil infiltration. Glandular loss of the glandular epithelium can be quantified by assessing the percentage of affected mucosa Erosion reflects loss of surface epithelium and is assessed by assessing the proportion of mucosa affected (e.g., mucosal bleeding). Mucosal thickness can be scored by assessing the overall thickness of the mucosa. The thickness increase can be evaluated by measuring the non-tangential area of ​​the part. Reflects glandular elongation and mucosal hyperplasia.

[0065] The overall histopathological score was calculated based on submucosal edema, inflammation, glandular defects, erosion, mucosal thickness, and hyperplasia. The score can be derived from one or more measurements of the formation of: A similar system is described in Example 1. Can be used on elephants.

[0066] In some embodiments, the inflammatory bowel disease is colitis, e.g., ulcerative colitis. Colitis may be accompanied by irritation, swelling, and other signs of inflammation in the colon. Ulcerative colitis causes tenderness and Ulcers are present.

[0067] In some embodiments, the inflammatory bowel disease is Crohn's disease, which is an inflammatory bowel disease limited to the colon. Crohn's disease can be localized to the colon and small intestine, but can also be present in other tissues, such as the small intestine. Inflammation of the mouth, anus, skin, eyes, joints, and / or liver may also be present. Yes.

[0068] In some embodiments, the subject has previously been treated with a TNF-α inhibitor alone and has In some embodiments, the subject has had a previous treatment for a chronic bowel disease that has not gone into remission. had been treated with IL-23 inhibitors alone and their inflammatory bowel disease had not gone into remission after previous treatment. The methods described herein can be used in combination with TNFα inhibitors (e.g., anti-TNFα antibodies) or IL- Treatment with either IL-23p19 inhibitors (e.g., anti-IL-23p19 antibodies) as monotherapy Anti-TNFα and anti-IL-23p19 antibodies may be beneficial for non-responders. The colon histopathology was significantly improved when both antibodies were administered (compared to either antibody alone). Based on the results described herein, which show substantial improvement in anti-IL-23p19 antibodies) and IL-23 inhibitors (e.g., anti-IL-23p19 antibodies) The body may respond much better to combinations with

[0069] In various embodiments, the IL-23 inhibitor is an anti-IL-23p19 antibody or its antigen. These include binding fragments that are capable of binding to the p19 subunit of IL-23.

[0070] In various embodiments, the TNFα inhibitor comprises an anti-TNFα antibody or antigen-binding fragment thereof. In some embodiments, the anti-IL-23p19 antibody is a human or humanized antibody. In some embodiments, the anti-TNFα antibody comprises a human antibody or a humanized antibody.

[0071] The anti-IL-23 and / or anti-TNFα antibodies may also be humanized or targeted to the antigen. Genetically engineered human antibodies while maintaining high affinity and other favorable biological properties. Humanized (or human) antibodies can optionally be prepared by combining the parental and humanized sequences. The process involves analyzing the parent sequence and various theoretical humanized products using a three-dimensional model of the sequence. Three-dimensional immunoglobulin models are commonly available and can be prepared by the skilled artisan. The likely three-dimensional conformation of the selected immunoglobulin sequence candidate is Computer programs are available to illustrate and display body structures. By comparing the residues, the likely role of the residues in the function of the candidate immunoglobulin sequence can be determined. This allows for analysis of residues that affect the antigen-binding ability of immunoglobulin candidates. In this way, desirable antibody properties such as enhanced affinity for the target antigen are achieved. Select framework (FR) residues from the consensus and import sequences to and can be combined.

[0072] The humanization or engineering of the antibodies of the present invention can be carried out according to the methods described in Winter (Jones et al., Nature 321:522(1986), Riechmann et al.,Na ture 332:323 (1988), Verhoeyen et al. nce 239:1534(1988)), Sims et al., J. Immuno l.151:2296(1993), Chothia and Lesk, J. Mol. Biol.196:901(1987), Carter et al., Proc.Na tl.Acad.SCi.USA89:4285(1992), Presta e t al., J. Immunol. 151:2623 (1993), and U.S. Pat. No. 723,323, No. 5,976,862, No. 5,824,514, No. 5,8 No. 17,483, No. 5,814,476, No. 5,763,192, No. 5,72 No. 3,323, No. 5,766,886, No. 5,714,352, No. 6,204 ,023, No. 6,180,370, No. 5,693,762, No. 5,530, No. 101, No. 5,585,089, No. 5,225,539, No. 4,816,5 Any known method, such as, but not limited to, those described in US Pat. No. 67,877, is also possible. and U.S. Pat. No. 6,239,999, each of which is incorporated herein by reference in its entirety.

[0073] In another aspect, a method of reducing colon inflammation in a subject with inflammatory bowel disease is provided. The method comprises administering an IL-23 inhibitor in an amount effective to reduce the first concurrent inflammation. and administering a TNFα inhibitor in an amount effective to reduce a second concurrent inflammation. The method includes reducing inflammation in the subject's colon to a level comparable to that of a normal patient's colon. The amount effective to reduce the first concurrent inflammation and the amount effective to reduce the second concurrent inflammation are The amount effective to prevent or reduce inflammation may be the same or different from the amount effective to prevent or reduce inflammation. The severity of the disease can be measured by biological analysis, the degree of weight loss, and the degree of inflammation.

[0074] In some embodiments, the colon of a subject after administration of an IL-23 inhibitor and a TNFα inhibitor. In histopathological studies of tissue samples from Limited inflammation may be reflected by the presence of only one or two small foci, with mononuclear inflammatory cells ( Mononuclear inflammatory cells (MNIC) are lymphoid aggregates in the background mucosa. It's likely to be the body.

[0075] In some embodiments, the colon of a subject after administration of an IL-23 inhibitor and a TNFα inhibitor. In histopathological studies of tissue samples from the Minimal glandular loss may involve only a localized area of ​​one or two glandular losses.

[0076] In some embodiments, the colon of a subject after administration of an IL-23 inhibitor and a TNFα inhibitor. Histopathological studies of tissue samples from the area show minimal or normal erosion scores. Minimal erosion may involve only one or two small localized areas of mucosal erosion.

[0077] In some embodiments, the colon of a subject after administration of an IL-23 inhibitor and a TNFα inhibitor. In histopathological studies of tissue samples from Minimal or normal mucosal thickness is defined as a thickness of mucosa compared to normal mucosal tissue thickness. This may include less than a 25% increase in

[0078] In some embodiments, after administration of an IL-23 inhibitor and a TNFα inhibitor, colon tissue The pathology is nearly identical (or identical) to normal tissue. Histopathology includes submucosal edema, inflammation, and adenomatous ducts. Evaluated by measuring one or more of the following: lesions, erosions, mucosal thickness, and hyperplasia Any or all of these parameters can be measured and scored. An exemplary scoring system is described in Example 1.

[0079] In various embodiments, the IL-23 inhibitor is an anti-IL-23p19 antibody or its antigen. Exemplary anti-IL-23p19 antibodies and fragments are listed in the "Anti-IL-23p19 Antibody and IL-23p19 Antibody" section of ... No. 7,491,399, issued to In various embodiments, the TNFα inhibitor is an anti-TNFα antibody or is an antigen-binding fragment thereof.

[0080] In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody are mixed in a ratio of 1:2 to 2:2. :1 (w / w) ratio. This ratio represents the dose of one antibody in a patient on a mg / kg basis. The dose of the antibody administered to the patient and the dose of other antibodies administered to the patient in mg / kg can be calculated. In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody can be It is administered in a ratio of 15:1 to 400:1 (w / w). This ratio is based on the patient's mg / kg dose. Dosage of one antibody in a patient and the dose of the other antibody in the same patient in mg / kg It can be calculated from the quantity.

[0081] Anti-TNFα antibody and anti-IL-23p19 antibody were administered in a ratio of 1:2 to 2:1 (w / w). For example, a human patient) to treat IBD (e.g., colitis and clonitis) in the subject. In some embodiments, the anti-TNFα antibody and The ratio of the anti-IL-23p19 antibody is 1:2 to 1:1.8 (w / w). In this embodiment, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is 1:1.9 to 1:1. 7 (w / w). In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody In some embodiments, the ratio of anti-T The ratio of NFα antibody to anti-IL-23p19 antibody was 1:1.7 to 1:1.5 (w / w). In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is 1: In some embodiments, the ratio of anti-TNFα antibody to anti-I antibody is 1.6 to 1:1.4 (w / w). The ratio of the antibody to the L-23p19 antibody is 1:1.5 to 1:1.3 (w / w). In this embodiment, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is 1:1.4 to 1:1. 2 (w / w). In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody In some embodiments, the ratio of anti-T The ratio of NFα antibody to anti-IL-23p19 antibody is 1:1.2 to 1:1 (w / w). In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is 1:1. In some embodiments, the ratio of the anti-TNFα antibody to the anti-IL- The ratio of the 23p19 antibody to the 23p19 antibody is 1:1 to 1.2:1 (w / w). The ratio of anti-TNFα antibody to anti-IL-23p19 antibody was 1.1:1 to 1.3:1 (w / In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is In some embodiments, the ratio of the anti-TNFα antibody to the anti-TNFα antibody is 1.2:1 to 1.4:1 (w / w). The ratio of the antibody to the anti-IL-23p19 antibody is 1.3:1 to 1.5:1 (w / w). In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is 1.4:1 to In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23 The ratio of the antibody to the p19 antibody is 1.5:1 to 1.7:1 (w / w). The ratio of anti-TNFα antibody to anti-IL-23p19 antibody was 1.6:1 to 1.8:1 (w / In some embodiments, the ratio of anti-TNFα antibody to anti-IL-23p19 antibody is In some embodiments, the ratio of the anti-TNFα antibody to the anti-TNFα antibody is 1.7:1 to 1.9:1 (w / w). The ratio of the antibody to the anti-IL-23p19 antibody is 1.8:1 to 2:1 (w / w). In some embodiments, the ratio of the anti-IL-23p19 antibody to the anti-TNFα antibody is about 1:2, 1:1.8, , 1:1.5, 1:1.2, 1:1, 1.2:1, 1.5:1, 1.8:1 or 2:1 ( w / w).

[0082] The lowest active dose of an anti-IL-23p19 antibody was administered in combination with a higher dose of an anti-TNFα antibody. and administering the compound to a subject (e.g., a human patient) to treat inflammatory bowel disease (e.g., colitis and Crohn's disease). ) can prevent the onset of the disease. The ratio with a higher dose of antibody can range from 1:400 to 1:15 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:400 In some embodiments, the anti-IL-23p19 antibody and the anti-IL-23p19 antibody are The ratio of the antibody to the TNFα antibody is 1:370 to 1:320 (w / w). The ratio of anti-IL-23p19 antibody to anti-TNFα antibody was 1:350 to 1:300 (w In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody are The ratio is 1:300 to 1:250 (w / w). The ratio of 3p19 antibody to anti-TNFα antibody is 1:280 to 1:230 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:250 In some embodiments, the anti-IL-23p19 antibody and the anti-IL-23p19 antibody are The ratio of the antibody to the TNFα antibody is 1:220 to 1:170 (w / w). The ratio of anti-IL-23p19 antibody to anti-TNFα antibody was 1:170 to 1:120 (w In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody are The ratio is 1:150 to 1:100 (w / w). The ratio of 3p19 antibody to anti-TNFα antibody is 1:120 to 1:80 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:100 to In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFR1 antibody is 1:60 (w / w). The ratio of the Fα antibody to the Fα antibody is 1:80 to 1:40 (w / w). The ratio of IL-23p19 antibody to anti-TNFα antibody is 1:60 to 1:30 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:5 In some embodiments, the ratio of the anti-IL-23p19 antibody to the anti-IL-23p19 antibody is 0:0 to 1:25 (w / w). The ratio of the antibody to the TNFα antibody is 1:40 to 1:20 (w / w). The ratio of anti-IL-23p19 antibody to anti-TNFα antibody was 1:35 to 1:15 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is about 1:400, 1:300, 1:200, 1:150, 1:100, 1:75, 1:50, 1:25 or 1:15 (w / w).

[0083] In some embodiments, the anti-TNFα antibody and the anti-IL-23p19 antibody are in a ratio of 15:1 to In some embodiments, the anti-IL-23p antibody is administered in a ratio of 400:1 (w / w). 19 antibody or antigen-binding fragment thereof, and b) anti-TNFα antibody or antigen-binding fragment thereof, In some embodiments, the antibody is administered to a subject in need thereof. a) an anti-IL-α antibody or antigen-binding fragment thereof, and b) an anti-TNFα antibody or antigen-binding fragment thereof are administered sequentially. a) a 23p19 antibody or an antigen-binding fragment thereof, and b) an anti-TNFα antibody or an antigen-binding fragment thereof , 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, or within 4 days This may be done.

[0084] In some embodiments, the antibody comprises: a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) The combination with an anti-TNFα antibody or antigen-binding fragment is not recommended for patients who have previously been treated with an anti-TNFα antibody alone. and is effective in treating subjects who have not achieved significant remission of inflammatory bowel disease. In some embodiments, a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) an anti-IL-23p19 antibody or antigen-binding fragment thereof. The combination with a TNFα antibody or antigen-binding fragment was previously compared with an anti-IL-23p19 antibody alone. and is effective in treating subjects who have been previously treated and have not achieved significant remission of inflammatory bowel disease. be.

[0085] In another aspect, a method for treating inflammatory bowel disease in a human subject is provided, the method comprising: (a) 0.0005 to 0.002 mg / kg of an anti-IL-23p19 antibody or its antigen binding antibody (b) administering 0.020 to 0.125 mg / kg of an anti-TNFα antibody or and administering the antigen-binding fragment thereof. In various embodiments, the method comprises administering to a subject a subject in need of an inflammatory In some embodiments, the inflammatory bowel disease is colitis. In some embodiments, the inflammatory bowel disease is Crohn's disease. In one embodiment, the method includes inhibiting weight loss (e.g., weight loss associated with inflammatory bowel disease). It is effective in

[0086] (a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof, and (b) an anti-TNFα antibody or The antigen-binding fragments may be administered simultaneously, sequentially, or within one day of each other.

[0087] In various embodiments, 0.020 to 0.125 mg / kg of anti-TNFα antibody and 0 0.020-0.125 mg / kg of anti-IL-23p19 antibody in a subject (e.g., a human patient) to treat IBD (e.g., colitis and Crohn's disease) in a subject. The combination of 50 μg each of anti-TNFα and anti-IL-23p19 in mice Early results from evaluating the combination show that this combination is superior to the single agents at the same dose. This suggests that the treatment enhances protection from colitis. See Example 1. In some embodiments, 0.020 to 0.040 mg / kg of anti-TNFα antibody and and 0.020 to 0.040 mg / kg of an anti-IL-23p19 antibody is administered to a human subject. In some embodiments, 0.030 to 0.050 mg / kg of anti-TNFα antibody and 0. A dose of 0.030 to 0.050 mg / kg of anti-IL-23p19 antibody is administered to a human subject. In some embodiments, 0.040 to 0.060 mg / kg of anti-TNFα antibody and 0.040 ~0.060 mg / kg of anti-IL-23p19 antibody is administered to human subjects. In this embodiment, 0.050 to 0.070 mg / kg of anti-TNFα antibody and 0.050 to 0. 0.70 mg / kg of anti-IL-23p19 antibody is administered to a human subject. In this study, 0.060 to 0.080 mg / kg of anti-TNFα antibody and 0.060 to 0.080 In some embodiments, the human subject is administered 1 mg / kg of an anti-IL-23p19 antibody. 0.070-0.090 mg / kg anti-TNFα antibody and 0.070-0.090 mg / In some embodiments, 0.0 kg of an anti-IL-23p19 antibody is administered to a human subject. 80 to 0.100 mg / kg of anti-TNFα antibody and 0.080 to 0.100 mg / kg of The anti-IL-23p19 antibody is administered to a human subject. In some embodiments, the antibody is administered in an amount of 0.090 to 100 mg / kg. 0.110 mg / kg anti-TNFα antibody and 0.090 to 0.110 mg / kg anti-IL In some embodiments, the antibody is administered to a human subject in the range of 0.100 to 0.1 25 mg / kg anti-TNFα antibody and 0.100 to 0.125 mg / kg anti-IL-23 The p19 antibody is administered to a human subject.

[0088] In various embodiments, the anti-IL-23p19 antibody is administered to a subject (e.g., a human patient) every 24 hours. Administered once a day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, or once a week In various embodiments, the anti-TNFα antibody is administered to a subject (e.g., a human patient) daily. , once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, or once a week In some embodiments, both the anti-IL-23p19 antibody and the anti-TNFα antibody are A subject (e.g., a human patient) may be administered daily, once every two days, once every three days, once every four days, once every five days, It is administered once every six days or once a week.

[0089] The anti-IL-23p19 antibody and the anti-TNFα antibody are co-administered to a subject (e.g., a human patient). Alternatively, the anti-IL-23p19 antibody and the anti-TNFα antibody may be administered separately to the subject. When administered separately, each antibody may be administered 3 hours, 6 hours, 12 hours, 1 day, or It may be administered within 2, 3, or 4 days.

[0090] In some embodiments, the antibody comprises: a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) The combination with an anti-TNFα antibody or antigen-binding fragment is not recommended for patients who have previously been treated with an anti-TNFα antibody alone. and is effective in treating subjects who have not achieved significant remission of inflammatory bowel disease. In some embodiments, a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) an anti-IL-23p19 antibody or antigen-binding fragment thereof. The combination with a TNFα antibody or antigen-binding fragment was previously compared with an anti-IL-23p19 antibody alone. and is effective in treating subjects who have been previously treated and have not achieved significant remission of inflammatory bowel disease. be.

[0091] In another embodiment, the minimum active dose of an anti-IL-23p19 antibody is administered in combination with a higher dose of an anti-TN and administering it with an Fα antibody to treat inflammatory bowel disease ( For example, to prevent recurrence of ulcerative colitis, undifferentiated colitis, and / or Crohn's disease The ratio of the minimum active dose of anti-IL-23p19 to a higher dose of anti-TNFα antibody In some embodiments, the concentration of anti-I The ratio of L-23p19 antibody to anti-TNFα antibody was 1:400 to 1:350 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1: In some embodiments, the ratio of anti-IL-23p19 antibody to anti-IL-23p19 antibody is 1:370 to 1:320 (w / w). The ratio of the antibody to the anti-TNFα antibody is 1:350 to 1:300 (w / w). In this embodiment, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:300 to 1:25. 0 (w / w). In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody In some embodiments, the ratio of anti-I to anti-I is 1:280 to 1:230 (w / w). The ratio of L-23p19 antibody to anti-TNFα antibody was 1:250 to 1:200 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1: In some embodiments, the anti-IL-23p19 antibody The ratio of the antibody to the anti-TNFα antibody is 1:170 to 1:120 (w / w). In this embodiment, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:150 to 1:10. 0 (w / w). In some embodiments, the anti-IL-23p19 antibody and the anti-TNFα antibody In some embodiments, the ratio of anti-IL-1 to anti-IL-1 is 1:120 to 1:80 (w / w). The ratio of -23p19 antibody to anti-TNFα antibody is 1:100 to 1:60 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:80 In some embodiments, the ratio of anti-IL-23p19 antibody to anti-T The ratio of the antibody to the NFα antibody is 1:60 to 1:30 (w / w). The ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1:50 to 1:25 (w / w). In some embodiments, the ratio of anti-IL-23p19 antibody to anti-TNFα antibody is 1: In some embodiments, the ratio of anti-IL-23p19 antibody to IL-23p19 antibody is 1:40 to 1:20 (w / w). The ratio of the anti-TNFα antibody to the anti-TNFα antibody is 1:35 to 1:15 (w / w). The ratio of anti-IL-23p19 antibody to anti-TNFα antibody was approximately 1:400, 1:300, and 1:100. :200, 1:150, 1:100, 1:75, 1:50, 1:25, or 1:15(w / w).

[0092] In various embodiments, the anti-IL-23p19 antibody is administered daily, once every two days, or once every three days. The dose is administered once every four days, once every five days, once every six days, or once a week. Anti-TNFα antibodies are administered daily, once every two days, once every three days, once every four days, once every five days, or once every six days. In some embodiments, the anti-IL-23p19 antibody is administered once daily or once weekly. Both the anti-TNFα antibody and the anti-TNFα antibody are administered to a subject (e.g., a human patient) daily, once every two days, or once every three days. It is administered once every four days, once every five days, once every six days, or once a week.

[0093] The anti-IL-23p19 antibody and the anti-TNFα antibody may be co-administered. The IL-23p19 antibody and the anti-TNFα antibody may be administered separately.

[0094] Anti-TNFα antibody (500 μg / mouse) treatment was performed at the minimum active dose of anti-IL-23p19 antibody. Combined with these doses, the efficacy of these antibodies was significantly improved compared to either single antibody treatment at these doses. Therefore, it is possible to provide excellent effectiveness in preventing the onset of colitis. See 5. The efficacy of this combination therapy versus anti-TNFα or anti-IL-23p19 monotherapy. Analysis of colonic gene signatures revealed genes enriched in fibroblasts and extracellular matrix tissues. The unique set of genes regulated by the combined therapy, cell types and This novel discovery was supported by the combination of antibodies against TNFα and IL-23p19. Combination therapy has been shown to provide superior efficacy in the treatment of colitis and inflammatory bowel syndrome. Furthermore, combined therapy with antibodies against TNFα and IL-23p19 has been shown to be effective. may have synergistic effects through the regulation of specific gene networks involved in mucosal healing.

[0095] The data in Example 5 demonstrates the efficacy of combination therapy with antibodies against TNFα and IL-23p19. Treatment with either antibody as monotherapy provided superior protection against colitis The colitis may be acute colitis. Without being bound by theory, Although we do not wish to be limited to these, transcriptomics and gene network analysis are Identify both overlapping and distinct molecular effects versus monotherapy to enhance wound repair processes. We identified a unique set of genes affected by the combined treatment involved in the process. Taken together, these findings suggest that combination therapy with anti-TNFα and anti-IL-23p19 antibodies may These findings suggest that the two compounds may provide a synergistic effect in reducing intestinal inflammation. The synergistic effects may occur through targeting common inflammatory pathways. This could result from treating individual cell types involved in the pathogenesis of IBD by affecting genes.

[0096] In some embodiments, the antibody comprises: a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) The combination with an anti-TNFα antibody or antigen-binding fragment is not recommended for patients who have previously been treated with an anti-TNFα antibody alone. and is effective in treating subjects who have not achieved significant remission of inflammatory bowel disease. In some embodiments, a) an anti-IL-23p19 antibody or antigen-binding fragment thereof; and b) an anti-IL-23p19 antibody or antigen-binding fragment thereof. The combination with a TNFα antibody or antigen-binding fragment was previously compared with an anti-IL-23p19 antibody alone. and is effective in treating subjects who have been previously treated and have not achieved significant remission of inflammatory bowel disease. be.

[0097] formulation Each of the anti-TNFα antibody and the anti-IL-23 (e.g., anti-IL-23p19) antibody is The stable formulation may be in a stable formulation. The stable formulation may be in a pharmaceutically acceptable formulation. (e.g., anti-IL-23p19) antibodies in saline or a phosphate buffer containing a selected salt. Contains buffers, and preservatives suitable for pharmaceutical or veterinary use and multi-use preservative formulations. The preservative formulation may comprise a preservative solution and a formulation containing at least one known or in some cases at least one of phenol, m-cresol, p-cresol Benzyl alcohol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite, Phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (e.g. (e.g., hexahydrate), alkylparaben (methyl, ethyl, propyl, butyl, etc.), benzochloride dsalkonium, benzethonium chloride, sodium dehydroacetate and thimerosal, and mixtures thereof. For example, about 0.0015%, or any range, value, or fraction thereof, may be used. Non-limiting examples include preservative-free, about 0.1-2% 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%), approximately 0.0 0.1-0.5% thimerosal (e.g., 0.005, 0.01), approximately 0.001-2. 0% phenol (e.g., 0.05, 0.25, 0.28, 0.5, 0.9, 1.0% ), 0.0005 to 1.0% alkylparabens (e.g., 0.00075, 0.000 9, 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%).

[0098] The aqueous diluent may further comprise a pharmaceutically acceptable preservative. Preferred preservatives include: Phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkylparaben (methyl, ethyl, propyl, butyl, etc.), chloride Benzalkonium, benzethonium chloride, sodium dehydroacetate and thimerosal or The preservatives used in the formulation include those selected from the group consisting of: The concentration is sufficient to produce an antimicrobial effect. Such concentration is maintained by the preservative selected. and are readily determined by one of ordinary skill in the art.

[0099] Other excipients, such as isotonicity agents, buffers, antioxidants, and preservative enhancers, may be added to the diluent. An isotonic agent such as glycerin is commonly used at a known concentration. Alternatively, a physiologically tolerable buffer may be added to provide improved pH control. A range of about pH 4 to about pH 10, and preferably about pH 5 to about pH 9, and most preferably The pH range may be broad, such as from about 6.0 to about 8.0. The formulation has a pH of about 6.8 to about 7.8. Suitable buffers include phosphate buffers, most preferably phosphate buffers. Preferably, sodium phosphate, especially phosphate buffered saline , PBS).

[0100] Other additives, such as Tween 20 (polyoxyethylene (20) sorbitan monolaurate) rate), Tween 40 (Polyoxyethylene (20) sorbitan monopalmitate) , Tween 80 (Polyoxyethylene (20) sorbitan monooleate), Plur onic F68 (polyoxyethylene polyoxypropylene block copolymer), and and PEG (polyethylene glycol), or a pharmaceutically acceptable solubilizer. Poloxamer 20 or 80 or Poloxamer 184 or 188, Pluronic Nonionic surfactants such as Polyl®, other block copolymers mers and chelating agents such as EDTA and EGTA may be added to the formulation or composition to promote coagulation. These additives can be used in pumps or pumps to administer the formulation. This may be useful when using a pharmaceutical container. This may reduce any tendency of the antibodies to aggregate.

[0101] The formulations of the present invention contain at least one anti-IL-23 antibody or anti-TNFα antibody. The preparation may be carried out by a process comprising mixing the solution with a buffer solution, such as saline or The aqueous diluent may be a phosphate buffer containing a selected salt. Mixing of the IL-23 antibody and buffer is carried out using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a quantity of at least one antibody in water or a buffer is and combining with the desired buffer in a quantity of water sufficient to provide the desired concentrations of protein and buffer. Variations on this process will be recognized by those skilled in the art. For example, The order of addition of the components, whether or not additional additives are used, and the temperature and pH at which the formulation is prepared are all important considerations. The dosage concentration and means of administration used are factors that can be optimized.

[0102] A stable or preserved formulation containing one or both of an anti-IL-23 antibody and an anti-TNFα antibody is a second solution containing a preservative or buffer and excipients as a clear solution or in an aqueous diluent; A dual-use vial containing at least one lyophilized antibody vial to be reconstituted in the vial. It can be provided to patients as a single solution vial or as a vial requiring reconstitution. Each dual vial can be reused multiple times for single or multiple patient treatments. cycles, thus providing a more convenient treatment regimen than is currently available. provide.

[0103] For parenteral administration, the anti-IL-23 antibody or anti-TNFα antibody is administered in a medically acceptable parenteral Solutions, suspensions, emulsions, particles, powders, provided in combination with a vehicle or separately The formulation may be in the form of a liquid or a lyophilized powder. Examples of such solvents include water, saline, and the like. Water, Ringer's solution, dextrose solution, and about 1-10% human serum albumin. Non-aqueous solvents such as liposomes and fixed oils can also be used. The powder contains additives that maintain isotonicity and chemical stability (e.g., sodium chloride for isotonicity). For chemical stability, buffers and preservatives). The formulation is sterilized by known or suitable techniques.

[0104] Suitable pharmaceutical carriers are described in Remington's, a standard reference text in this field. Published in the latest edition of Pharmaceutical Sciences, A. Osol It has been done.

[0105] For administering a pharmaceutically effective amount of at least one anti-IL-23 antibody or anti-TNFα antibody. Many known and developed methods can be used in accordance with the present invention. Although pulmonary administration has been used in the description, other modes of administration may be used in accordance with the present invention with suitable results. The IL-23p19 antibody of the present invention may be in the form of a solution, emulsion, or concentrate in a carrier. as a globulin or suspension, or as a dry powder, by inhalation or as described herein Various devices suitable for administration by conventional or other methods known in the art are available. The delivery can be performed using any of the following devices and methods:

[0106] Formulations for parenteral administration may contain common excipients. Common excipients include: Examples of suitable solvents include, but are not limited to, sterile water or saline, and polyaniline such as polyethylene glycol. Examples of suitable injectable solutions include alkylene glycol, vegetable oils, and hydrogenated naphthalenes. Oily suspensions may be prepared in accordance with known methods using suitable emulsifying or wetting agents and suspending agents. Injectable preparations can be prepared, for example, by aqueous solutions, sterile injectable solutions, or suspensions in solvents. It may also be a non-toxic parenterally administrable diluent. , water, Ringer's solution, isotonic saline, etc. are possible, and the usual solvent or suspension solvent is For this purpose, any sterile, fixed oil may be employed, whether natural, synthetic or Semi-synthetic fatty oils or fatty acids, natural or synthetic or semi-synthetic monoglycerides or diglycerides Any type of fixed oil and fatty acid may be used, including glycerides or triglycerides. This can be done.

[0107] Formulations for oral administration include the use of adjuvants to artificially increase the permeability of the intestinal wall. (e.g., resorcinol, and polyoxyethylene oleyl ether and n-hexyl Simultaneous administration of a non-ionic surfactant such as methyl methyl ethylene ether, and an enzyme Enzyme inhibitors to inhibit catalytic degradation (e.g., pancreatic trypsin inhibitor, diisopropyl fluoride) Oral administration, such as simultaneous administration of orofacial acid (DFF) and trasylol. Drugs intended for oral, mucosal, nasal, pulmonary, transvaginal, or rectal administration Delivery of hydrophilic agents including proteins and antibodies and combinations of at least two surfactants Formulations for oral administration are taught in U.S. Patent No. 6,309,663. The active ingredient compounds in the dosage form are sucrose, lactose, cellulose, mannitol, trehalose, and lactic acid bacteria. Phenose, maltitol, dextran, starch, agar, alginate, chitin, chitin Tosan, pectin, tragacanth gum, gum arabic, gelatin, collagen, casein , albumin, synthetic or semi-synthetic polymers, and glycerides. These dosage forms can also be mixed with other types of additives, such as inert diluents. Additives, lubricants (magnesium stearate, etc.), parabens, preservatives (sorbic acid, ascorbic acid, corbic acid, α-tocopherol, etc.), antioxidants (cysteine, etc.), disintegrants, binders , thickeners, buffers, sweeteners, flavoring agents, perfumes, and the like.

[0108] The compounds of the present invention can be administered to subjects in a single dose over an extended period of time, for example, from one week to one year. It may be desirable to deliver the drug to a patient in a variety of sustained release, depot, or implantable dosage forms. For example, the dosage form may be a pharmaceutically acceptable carrier for a compound that has low solubility in body fluids. Non-toxic salts, such as (a) phosphoric acid, sulfuric acid, citric acid, tartaric acid, tannic acid, pamoic acid, arachidic acid, acid, polyglutamic acid, naphthalene mono- or disulfonic acid, polygalacturonic acid, etc. (b) Acid addition salts with any polybasic acid, (c) zinc, calcium, bismuth, barium, magnesium polyvalent metal cations such as aluminum, copper, cobalt, nickel, cadmium, or , for example, formed from N,N'-dibenzyl-ethylenediamine or ethylenediamine or (c) a combination of (a) and (b), e.g., a salt having an organic cation In addition, the compounds of the present invention, or preferably those described above, may contain zinc tannate. A relatively insoluble salt such as monostearate may be dissolved in a suitable solution for injection, e.g., sesame oil. They can be formulated in gels such as aluminum phosphate gel. Particularly preferred salts are , zinc salt, zinc tannate, pamoate, etc. [Example]

[0109] The present invention is also illustrated and demonstrated by the following examples. The use of these and other examples anywhere in the present specification is merely illustrative and does not constitute a limitation on the scope of the present invention. or limiting the scope and meaning of any exemplary term. Likewise, the present invention does not encompass any particular preferred embodiment described herein. Indeed, many modifications and variations of the invention will become apparent to those skilled in the art upon reading this specification. Such variations may be apparent to those skilled in the art and do not depart from the invention in spirit or scope. The present invention is therefore defined by the appended claims and the appended claims. This patent is limited only by the full scope of equivalents to which the patent is entitled.

[0110] Example 1: Dose ranging of monotherapy with antibodies to TNFα or IL-23p19 and combination studies in a CD40 antibody-induced colitis model Three separate studies were conducted. In all three studies, animals were randomized by body weight. Each animal was assigned to a treatment group, labeled with a specific number from 1 to 10. by intraperitoneal injection of 0.2 mg of CD40 agonist antibody in 0.2 mL of PBS The day before disease induction (day 0), the solution was administered as a single intraperitoneal (ip) injection. Vehicle (PBS) and mAb treatment were administered (day -1).

[0111] Naive control mice were not treated and were kept in separate cages until termination on day 7. Observations for clinical signs were made daily. Body weights were measured and the animals were then fed daily from day -1 until termination on day 7. At the end of the study (day 7), animals were euthanized by CO2 overdose and histologically analyzed. Colon tissue was removed and processed appropriately for analysis.

[0112] After euthanasia, the colon, defined as the intestinal section between the cecum and rectum, was excised and washed with ice-cold PBS. The fecal contents were removed by flushing. One centimeter of the proximal colon was placed in a histology cassette. The cassettes were then immersed in fixative (10% neutral buffered formalin, NBF). After 24 hours, the cassettes were fixed. The remaining colon tissue was removed from the solution, transferred to 70% ethanol, and stored refrigerated until processing. Divide into two equal parts, and snap-frozen the first third in liquid nitrogen for PK analysis. One third was frozen in liquid nitrogen for cytokine analysis. The last third (distal, closest to the rectum) is removed after all animals have been euthanized and the tissue properly removed. The 1 mL RNA was then stored in a 1 mL reservoir until frozen for RNA extraction and gene expression analysis. All frozen samples were stored on ice in an RNAlater (Ambion™). was stored at −80° C. until further processing.

[0113] In all three studies, animals were randomized by weight and assigned to treatment groups. Each animal was labeled with a specific number from 1 to 10. 0.2 mg of the antibody was added to 0.2 mL of PBS. The day before (day 0), induce disease by intraperitoneal injection of CD40 agonist antibody. Vehicle (PBS) and mAb treatments were administered as a single intraperitoneal (ip) injection (day -1). Naive control mice were not treated and were kept in separate cages until termination on day 7. Observations for clinical signs of disease were conducted daily. Body weights were measured and the animals were fed every day from day -1 until termination on day 7. Animals were euthanized on day 7 by CO2 overdose and colon tissue was analyzed histologically. It was removed and processed appropriately for analysis.

[0114] In the first study (Study 1), anti-TNFα mAb or anti-IL-23p19 mAb was administered to These antibodies were evaluated in a CD40 colitis model. Individually at a dose of 50 μg or in combination (i.e., 500 μg + 500 μg / each The protocol is summarized in Table 1 below.

[0115] [Table 2]

[0116] CNTO3723 is a murine anti-IL-23p19 monoclonal antibody (IL-2 CNTO5048 is a murine anti-TNFα monoclonal antibody. CNTO6601 was used throughout the experiment. Refers to the isotype control used. CNTO6601 inhibits TNFα or IL-23p1 9.

[0117] The anti-inflammatory activity of anti-TNFα and anti-IL-23p19 antibody treatment (alone or in combination) was evaluated. The effect of agonist antibody-mediated costimulation was evaluated in an anti-CD40 antibody-induced colitis model. Ligation of CD40 inhibits lymphopenic (T and B cell deficient) RAG2 - / - It induces acute innate systemic and colonic inflammatory responses in mice, and the colonic inflammatory response is It peaks on day 7 and then recovers (ELN Immunopharmacolo gy WC-2015-00008). IL-23 inhibits local colonic Drives inflammation.

[0118] Expression of TNFα regulates the development of systemic diseases (e.g., weight loss), but TNFα is largely (1) The present inventors have demonstrated that the effects of IFN-γ on intestinal gene expression are minimal. To investigate the different molecular effects of anti-TNFα antibody treatment and anti-IL-23p19 antibody treatment, and to Combination treatment with Fα and anti-IL-23p19 demonstrated greater efficacy than either monotherapy On day 1, RAG2 - / - Mice were given 0.5 mg or 0.05 mg of anti-TNFα antibody (CNTO5048), 0.5 mg or 0.05 mg Anti-IL-23p19 antibody (CNTO3732), both antibodies (0.5 mg or 0 0.05mg), 1.0mg of isotype control antibody (CNTO6601), or 10mL PBS was administered intraperitoneally once at a dose of 1 / kg. (RA used in all examples herein) G2 - / - Mice were 8-10 week old female mice supplied by Taconic Farms. One day later, on day 0, all animals were intraperitoneally injected with anti-CD40 antibody (0.2 mg). A challenge was performed to induce inflammation.

[0119] Weight loss analysis was performed after treatment with low dose (50 μg) and high dose (500 μg) of antibody. Body weights were measured from day 1, when the mice were injected with antibody or PBS, until termination on day 7. , monitored.

[0120] Exemplary data are shown in Figures 1A and 1B. Each line has an error bar for the standard error (n = 1). 9 antibody treatment, n=5 PBS control, n=3 naive control), - Shown as percentage change from day 1 (dotted line). Some error bars are within the symbol size. Figure 1A shows low-dose antibody treatment (50 μg / mouse), and Figure 1 B shows high-dose antibody treatment (500 μg / mouse). The statistical significance of the difference in weight loss between the control group and the control group was determined using Dunnett's multiple comparison test. The results were analyzed by two-way ANOVA, and the P values ​​for each time point are shown in the table. P values ​​indicating significance are in bold. Emphasis in italics. ELN:Immunopharmacology WC- 2018-00034, Immunopharmacology WC-2018-00 033.

[0121] The CD40 mAb-induced colitis model was performed 24–48 h after administration of CD40 agonist antibodies. An initial rapid weight loss within 2 hours, followed by recovery, and a second weight loss period on days 5-7. The anti-IL-23p19 antibody (0.5 mg and 0.0 mg) was used in combination with the anti-IL-23p19 antibody (0.5 mg and 0.0 mg). Single treatment with 5 mg of riboflavin did not protect mice from the initial rapid weight loss, but promotes faster recovery from the second stage of the disease, as shown in Figures 1A and 1B. Overall, protection against weight loss was dose-dependent and partial.

[0122] In contrast, monotherapy with anti-TNFα antibodies (0.5 mg and 0.05 mg) resulted in both The dose completely protected the mice from weight loss for the entire duration of the study. Similar to the single antibody treatment for HIV, the combination treatment demonstrated significant improvements in weight loss at both doses. Low doses of anti-TNFα / IL-23p19 resulted in complete protection (Figures 1A and 1B). No side effects were observed with either or high dose combination therapy.

[0123] At termination (day 7), colon histopathological scores were obtained for the low-dose and high-dose antibody treatment groups. The proximal colonic segments were stained with H&E and analyzed by a blinded pathologist according to the following protocol: Therefore, a severity score of 0 to 20 was used to examine histopathological changes.

[0124] For the proximal colon, two small pieces were cut and embedded in paraffin. Sections (5 μm) were Two colon segments from each animal were sectioned and stained with hematoxylin and eosin (H&E). The specimens were individually evaluated for histopathology and the mean values ​​per animal were used for group analysis. Each color segment represents submucosal edema within the non-tangential area that is thought to best represent the severity of this change. Quantification was performed by measuring the thickness from the muscularis mucosae to the inner border of the superficial muscularis.

[0125] The inflammation score reflects the degree of macrophage, lymphocyte, and neutrophil (PMN) infiltration. Severity scores were assigned according to the following criteria: 0=normal; 0.5 = minimal; one or two small foci, mononuclear inflammatory cells (MNIC) in mucosal linings However, the aggregates are likely to be lymphoid aggregates of the background. If , the ensemble is not scored as anomalous. 1 = Minimal, MNIC and neutrophils or larger focal areas with minimal diffusion, glandular separation There may be no, mostly submucosal edema or in the mesenteric region. 2 = Mild, mildly diffuse, or multiple, affecting 11-25% of the mucosa; With focal or multiple glandular separation, but no separation in most areas 3 = Moderate, 26-50% of the mucosa with minimal to mild focal or multiple inflammatory cell infiltration The remaining areas of the mucosa are affected by the partial glandular separation, with some areas being inflamed and others being milder. No glandular separation due to 4 = marked, 51-75% of the mucosa affected by mild to moderate glandular separation due to inflammatory cell infiltration affected, while the remaining areas of the mucosa are minimal to mild. All glands have some separation due to infiltration 5 = Severe, 76-100% of the mucosa with moderate to marked areas of glandular separation due to inflammatory cell infiltration affected, the remaining areas of the mucosa are mild to moderate

[0126] A glandular loss score was determined. The loss of crypt epithelium and remaining glandular epithelium was measured in most of the affected mucosa. Based on this percentage, the score is as follows: 0=none 0.5 = minimal, one or two small focal glandular defects or areas of mucosal erosion 1 = minimal, 1–10% of the mucosa affected 2 = mild, 11–25% of the mucosa affected 3 = moderate, 26-50% of the mucosa affected 4 = marked, 51-75% of the mucosa affected 5 = severe, 76-100% of the mucosa is affected

[0127] An erosion score was determined. The loss of surface epithelium was calculated based on the approximate percentage of mucosa affected. This was generally scored as follows: mucosal bleeding (clinical and postmortem) associated with bleeding (reflecting the bleeding seen in 0=none 0.5 = minimal, one or two small focal glandular defects or areas of mucosal erosion 1 = minimal, 1–10% of the mucosa affected 2 = mild, 11–25% of the mucosa affected 3 = moderate, 26-50% of the mucosa affected 4 = marked, 51-75% of the mucosa affected 5 = severe, 76-100% of the mucosa is affected

[0128] Mucosal thickness and hyperplasia scores were determined. Mucosal thickness was the thickness of the area that best represents the thickness of the entire mucosa. The non-tangential area was measured. This parameter indicates glandular elongation and mucosal hyperplasia. A is derived from measurements as follows: 0=≦200μm=normal 0.5 = 201~250μm = minimum 1=251~350μm=min. 2 = 351-450 μm = mild 3=451~550μm=moderate 4 = 551~650μm = significant 5=>650μm=severe

[0129] The histopathological score is the sum of the inflammation, glandular loss, erosion, and hyperplasia scores. The histopathological scores range from 0 to 20. Figures 2A and 2B show the histopathological scores. Bars represent group means with standard errors. In naive animals, histopathological findings were observed. Figure 2A shows the results for a low dose of antibody (50 μg / mouse). The results of the dose treatment group (500 μg / mouse) are shown. Treatment groups, vehicle control, and isotype Differences from the control were assessed for significance using one-way ANOVA and Sidak's multiple comparison test. Analysis: ELN:Immunopharmacology WC-2018-0003 4, Immunopharmacology WC-2018-00033.

[0130] In the proximal colon, treatment with isotype antibody (1000 μg / mouse) significantly reduced the incidence of disease-related leukemia ( showed a trend toward a decrease in histopathology when compared with PBS, but this was statistically did not reach significance. Anti-TNFα antibody monotherapy significantly reduced the risk of gliomas compared with isotype control. When administered at a high dose (500 μg, Figure 2B), colonic inflammation was significantly reduced, whereas a low dose (5 0 μg, Fig. 2A) did not significantly reduce the

[0131] A single administration of anti-IL-23p19 antibody was highly effective at a high dose (500 μg, Figure 2B). At a low dose (50 μg, Figure 2A), monotherapy was effective and completely prevented the onset of colitis. The treatment showed a significant reduction in histopathology compared with the isotype group, but did not completely abrogate colitis. The combination of high doses of both antibodies (500 μg anti-TNFα + 500 μg Anti-IL-23p19 / mouse (Fig. 2B) was similar to high-dose anti-IL-23p19 monotherapy. completely prevented colitis in the disease model.

[0132] Low-dose combination treatment (50 μg anti-TNFα + 50 μg anti-IL-23p19 / mouse) , Figure 2A) was significantly more effective than single-agent anti-TNFα treatment and inhibited IL-23p19. showed a trend towards improved protection compared to monotherapy against The possibility of sexuality was demonstrated.

[0133] Example 2: Anti-TNFα and anti-IL-23p19 treatments affect intrinsic genes in the gut influence Anti-TNFα and anti-IL-23p19 treatments provide readouts of systemic and local inflammation This example shows the differential effect of the treatment in Example 1 above on intestinal gene expression. To generate a gut gene signature, we evaluated whether the different molecular effects of , mRNA was isolated from the distal colon and submitted for microarray analysis.

[0134] For RNA extraction, thaw tissue samples on ice and add 900 μL of Qiazol (Qiagen). en) and transferred to a new tube containing one metal bead, followed by tissue disruption and homogenization. For the purification, a TissueLyser II was used, and the vibration frequency was 30 s. -1 In 1 Each sample was dissolved by stirring for 1 minute. 180 μL of chloroform was added to each sample. Add, vortex for 30 seconds, incubate at room temperature for 2 minutes, and then rotate at 14,000 rpm. The mixture was separated into organic and aqueous phases by centrifugation at 4°C for 15 minutes. The RNeasy 96-well plate kit (Qiagen) was used, which includes a single digestion step. RNA extraction was performed using 150 μL of aqueous phase, all following the manufacturer's protocol. The quality and quantity of isolated RNA was confirmed using Nanodrop® according to the manufacturer's protocol. by Nanodrop on the 8000 instrument (ThermoScientific) and Caliper instrument (Life Science) with LabChip GX (GX DNA 5K / RNA for use with Touch / GXII Touch HT For caliper analysis, colonic RNA aliquots were collected and analyzed by the Caliper CZE Chip. The solution was diluted 1:4 with molecular grade water.

[0135] The following exclusion criteria were used to determine which samples were eligible for gene expression analysis by microarray: The ratio of Nanodrop absorbance 260 / 280 (times relative to nucleic acid) was determined to be acceptable. The absorbance ratio of Nanodrop 260 / 2 The ratio of the absorbance of the Nanodrop to the amount of salt (amount of nucleic acid) should be close to 2. If the RIN was less than 1.5, repurification was performed. The RNA integrity number (RI) should be between 5 and 10. If the score is less than 5, it may be affected by the accuracy of the microarray analysis. For ELISA analysis, RNA was transferred to BioStorage Technologies (India). The product was shipped to Ianapolis, IN.

[0136] To compare the effects of anti-TNFα or anti-IL-23p19 with those of isotype control treatments Differential gene expression analysis was performed using 50 μg anti-IL-23p19 dose or 5 μg anti-IL-23p19 dose. Treatment with either the 0.001 mg or 0.001 mg anti-TNFα doses resulted in similar levels of histological inflammation. This resulted in a decrease in the expression of cellular invasive genes (Fig. 2). To mitigate potential confounding effects, we selected these colonic gene expression signatures for further evaluation. Select.

[0137] The mouse gene signature for each treatment was analyzed for overlap and enrichment in biological pathways. The data were evaluated (Enrichr: http: / / amp.pharm.mssm.edu / Enrichr / ). Individual genes generated from anti-TNFα or anti-IL-23p19 treatment. The overlap of the child signatures was relatively small, with only 11% of genes shared between the signatures. No specific pathway enrichment was observed. Gene signatures for anti-TNFα therapy (26 7 genes (FDR < 0.05, FC > 1.2) were identified as metabolic pathways and cytokines. The anti-IL-23p19 gene signature ( 765 genes (FDR < 0.05, FC > 1.2) were associated with circadian rhythm and p53 signaling. Enriched in transmission.

[0138] Example 3: Single antibody therapy with anti-TNFα and anti-IL-23p19 improves the efficacy of human IBD Affects overlapping and separate parts of the network Mount Sinai School of Medicine (New York) In collaboration with the CERTIFI Crohn's Disease Clinical Trial (847 IBD biopsies, 28 non-IBD biopsies), Transcriptional data from intestinal biopsy samples collected in IBD control biopsies (7,796 gene nodes) To integrate the genetic data, a Bayesian network predictive model was generated. (7 ,10) This type of molecular integrated network is a gene-gene interaction in disease association. We provide a data-driven framework for studying interactions in a mouse colitis model. The generated anti-TNFα and anti-IL-23p19 monotherapy gene signatures were compared with clinical disease. Integrating mouse gene signatures with human IBD patient gene networks to translate As above, a 50 μg anti-IL-23p19 dose and a 500 μg anti-TNFα dose were used. These were selected for evaluation based on their similar effects on histological inflammation. Ta.

[0139] To bridge mouse model data to the human IBD network, we first Genes were compared with their human orthologues (767 genes for anti-IL-23p19 and anti-TNFα By mapping the gene signatures to the 274 genes in the current study, we were able to identify the gene signatures for each treatment. A "humanized" version was generated. The mouse gene was then cloned using NCBI HomoloGene ( https: / / www.ncbi.nlm.nih.gov / homologene) Database (Build 68, 04 / 14 / 2014) was used to map to human orthologs. Each NCBI gene ID for each profiled mouse gene was mapped to All corresponding human members of the same cluster of putative orthologs were matched.

[0140] The one-sided Fisher's exact test E value (Bonferroni corrected P value) is 0 Database terms were considered significant if they were less than .05.

[0141] To determine gene enrichment of IBD GWAS loci within gene subnetworks Then, a hypergeometric distribution test was performed in Excel (HYPGEOM.DIST function). The gene list used to enrich for GWAS loci in BD was published by Jostins et al. l,Nature 2012(8)and Liu et al,Nature Gen Retrieved from etics 2015(9).

[0142] These humanized gene signatures were used to perform enrichment analysis of individual therapeutic signatures in humans. The gene signature for anti-TNFα therapy was expanded to include responses to stress and lipids. Cellular responses, reactive oxygen species metabolism, inflammatory response genes, and down-regulated genes in patient biopsies Anti-IL-23p19 therapeutic signatures were enriched in genes involved in the regulation of cell metabolism, proliferation, and , and were enriched in genes downregulated in IBD patient biopsies.

[0143] These humanized gene signatures were then used to rank the CERTIFI Bayesian network. and treatment subnets using a web-based network visualization tool. The gene list was generated as a tab-delimited file and imported. The gene list was analyzed using the T26 Pan-Intestine Bayesian Net work (CERTIFI network (7)) and their first neighboring genes ( Genes within one step of the selected gene, either incoming or outgoing You created a subnetwork using either

[0144] These therapeutic subnetworks were designed to target anti-TNFα or anti-IL-23 antibodies in mouse models. Genes and networks altered by p19 treatment and reflected in human IBD tissues Therefore, the enrichment solution of these sub-networks The analysis will identify the biological pathways targeted by each treatment in the context of human disease tissue. can provide insight into the

[0145] Figures 3A and 3B are projected onto the CERTIFI human IBD gene expression network. Furthermore, anti-TNFα or anti-IL-23p19 monotherapy from an anti-CD40 model of murine colitis First neighbors of genes in the human IBD network Therapeutic sub-networks were generated by extracting the anti-TNFα and anti-IL The overlap between genes present within the -23p19 subnetwork is shown by the central Venn diagram. The largest connected component of the shared subnetwork of anti-TNFα and anti-IL-23p19 is shown in Figure 3B.

[0146] In the analysis of the intersection of the original gene signatures, specific biologies are enriched. Although not shown, a network shared by both anti-TNFα and anti-IL-23p19 was IBD is performed by centralizing the largest connected components of the network neighborhood. Enrichment of genes dysregulated in patient tissues as well as genes from IBD GWAS loci The effectiveness of these different mechanisms has been revealed through targeting shared core inflammatory pathways. These findings suggest that the interaction between these two treatment sub-networks may be partially mediated by the The difference was observed in the IBD GWAS locus (p=0.001) and in IBD patient tissue. Significantly enriched in genes significantly regulated by multiple signatures (E of top signatures) The unique part of the anti-TNF subnetwork is composed of neutrophils and C D11b +Highly enriched in the macrophage gene signature (E value 8.0, respectively) .28e-10 and 2.41e-06), while the anti-IL-23p19 subnetwork The intrinsic portion was highly enriched in colonic epithelial cells (E value 1.27e-32), which indicates that Cytokines such as IL-17A and IL-22 that affect the biology of epithelial cells This is consistent with the role of IL-23 in promoting the expression of anti-TNFα and anti-IL- The relative enrichment in myeloid and epithelial cells within the 23p19 unique region was Combination antibody therapy may improve outcomes by targeting distinct cell types involved in IBD pathogenesis. This led to the further hypothesis that orthogonal mouse models of intestinal inflammation could provide benefits. A T cell transfer model of colitis (ELN:jperrigo-2016-0000 2) Gene signatures derived from anti-TNFα or anti-IL-23p19 therapeutic treatment were analyzed. Very similar results were observed when the same type of network analysis was performed using Taken together, these network analyses demonstrate the effects of anti-TNFα and anti-IL-23p19. Although the mechanisms differ, they suggest convergence on the molecular drivers of intestinal inflammation.

[0147] Example 4: Anti-TNFα and anti-IL-23p19 antibodies in anti-CD40 antibody-induced colitis Supplementary analysis of dose ranges for systemic therapy (Study 2) To further evaluate the efficacy of the combination therapy, we performed a CD40 antibody-induced colitis model. An extended dose-response study was performed to determine the minimal effective dose of each antibody. The day before disease induction with 40 agonist antibodies, female RAG2 mice were - / - Anti-IL-23p in mice 19 antibodies (CNTO3723, 50, 15, 5, 1.5, 0.5, 0.15 μg / mouse) ), anti-TNFα antibody (CNTO5048, 150 and 15 μg / mouse), or isotonic acid A control type (50 μg / mouse) was administered intraperitoneally. The protocol is summarized in Table 2 below.

[0148] [Table 3]

[0149] Mice were injected with antibody or PBS and their body weights were monitored from day 1 until termination on day 7. The data are shown in Figures 4A-4D. Each line represents the standard error (n = 10 antibody treatments, n = 5 represents the group mean with PBS control, n=3 naive control, and changes from day -1 (dotted line). The significance of differences relative to the isotype control group was determined by Dunnett's multiple regression analysis. Results for each treatment group were analyzed by two-way ANOVA using a comparison test, and the results for each study day were The p-values ​​obtained are shown in the table. Significant p-values ​​are highlighted in bold / italics. Ru. ELN: Immunopharmacology WC-2016-00038, I mmunopharmacology WC-2018-00033.

[0150] There was a partially significant increase in body weight loss in the isotype control group compared to the vehicle control. Treatment with anti-IL-23p19 antibodies was observed at two higher doses (15, 50 μg). g / mouse), showed partial dose-dependent protection against weight loss from day 2. As shown in 4B, only at the lowest dose of anti-IL-23p19 antibody (0.15 μg / mouse) However, no protection from weight loss was observed. Treatment with anti-TNFα antibodies was observed at higher doses. (150 μg / mouse) completely protected against weight loss, whereas lower doses (1 5 μg / mouse) only provided partial protection. See Figure 4C.

[0151] Following single antibody treatment for dose ranging, histopathological analysis of the proximal colon was performed as follows: At termination (day 7), proximal colon segments were removed, flushed, fixed, and then analyzed by H&E. The histopathological changes of the stained samples were analyzed using the protocol in Example 1 above. The data were analyzed by a blinded pathologist using a severity score of 0 to 20. These results are shown in Figures 5A to 5C. No histopathological findings were observed in naive animals. Differences between antibody-treated groups and their respective isotype controls were analyzed by one-way ANOVA and Sidak's method. Significance was analyzed by multiple comparison tests. Lines indicate group medians. ELN:Immu nopharmacology WC-2016-00038, Immunopharm acology WC-2018-00033.

[0152] Colon histopathology showed significant improvement with anti-IL-23p19 antibody treatment, as shown in Figure 5B. At a dose of 50 μg / mouse, anti-IL-23p19 demonstrated dose-dependent protection from intestinal inflammation. Antibody treatment provided near complete protection. Antibody doses of 15 μg and 5 μg provided partial protection. Protection was not observed at doses below 1.5 μg. At two dose levels (150 and 15 μg), no significant therapeutic effect was observed in colon histopathology. No effect was detected (see Figure 5C). These results suggest that IL-23 signaling This confirms that blocking transmission is highly effective against colitis in this model. Inhibition of TNFα is effective against systemic inflammation (as measured by improved weight loss). (when administered intravenously) provides only modest protection against colitis in this model.

[0153] Example 5: Comparison of a fixed dose of anti-TNFα antibody with various doses of anti-TNFα antibody in a CD40 colitis model Determining the anti-inflammatory activity of combination with IL-23p19 antibody (Study 3) A fixed dose of anti-TNFα antibody (500 μg / mouse) was administered in combination with various doses of anti-IL-23p1 By using it in combination with 9 antibodies (1.5, 5, 25 μg / mouse), A study on the effect of anti-IL-23p19 antibodies on the CD40-induced colitis was performed in a CD40-induced colitis model. The protocol is summarized in Table 3 below.

[0154] [Table 4]

[0155] Monotherapy with high-dose anti-TNFα and low-dose anti-IL-23p19 antibodies and their Assays for weight loss after combination treatment were performed as follows.

[0156] Mice were injected with antibodies (isotype control: 525 μg, anti-TNFα: 500 μg, anti-IL-2 3p19:25, 5, 1.5 μg) or PBS (10 mL / kg) was injected on day 1. Body weights were monitored from day 1 until termination on day 7. Data are shown in Figure 6. Each line represents the group mean. (n=10 antibody-treated and vehicle, n=5 naive control), change from day -1 (dotted line). The significance of differences relative to the isotype control group was determined by Dunnett's multiple regression analysis. Each treatment group was analyzed by two-way ANOVA using the comparison test. p for each study day Values ​​are shown in tables and, where significant, are highlighted in bold / italics. ELN:Imm unopharmacology WC-2016-00066, Immunophar macology WC-2018-00033.

[0157] Consistent with previous studies, as shown in Figure 6B, high-dose anti-TNFα antibody had no effect on weight loss. In contrast, monotherapy with anti-IL-23p19 antibody provided complete protection at all doses. provided partial protection from weight loss, especially during the later stages of anti-CD40 antibody-induced disease See Figure 6C. The combination of anti-TNFα and anti-IL-23p19 antibodies Compared to monotherapy, it did not provide any further detectable effect on inhibiting weight loss ( Without wishing to be bound by theory, this effect is thought to be due to the anti-TNFα antibody. This may be due to the strong effect of monotherapy on this parameter.

[0158] Single-agent and combination anti-TNFα and low-dose anti-IL-23p19 antibodies After the treatment, histopathological analysis of the proximal colon was performed. At the end of the treatment period (day 7), the proximal colon tissue samples were The tissue was removed, rinsed, fixed, and then stained with H&E as described in Example 1 above. Histopathological changes were assessed by a blinded pathologist using a severity score of 0 to 20. The data are shown in Figures 7A to 7C. In naive animals, the histopathological findings were No differences were observed between the antibody-treated groups and their respective isotype controls using one-way ANOVA. Significance was analyzed by VA and Sidak's multiple comparison test. Lines indicate group medians. . ELN: Immunopharmacology WC-2016-00066, Im munopharmacology WC-2018-00033.

[0159] As shown in Figures 7A to 7C, the anti-TNFα antibody (500 μg / mouse) Anti-IL-2 did not provide significant protection in colon histopathology compared with control. Treatment with 3p19 antibody (1.5, 5, and 25 μg / mouse) provided dose-dependent protection from colitis. The lowest dose (1.5 μg / mouse) did not provide protection. At these doses (5 and 25 μg / mouse), partial protection from colitis was observed.

[0160] Because the amount of antibody used for anti-IL-23p19 is low, single-agent anti-IL-23p19 treatment Statistical significance was calculated relative to the vehicle control, but not the high-dose (525 μg / mouse) eye. All combination treatments were compared with single-agent anti-T cells. Compared to NFα treatment, it showed significant protection from colonic inflammation. See Figures 7A-7C. Of note, the low combination dose evaluated (500 μg / mouse TNFα + 1 0.5μg / mouse anti-IL-23p19), colon histopathology revealed that both monotherapy Treatment with IFN-γ failed to provide protection, but when given in combination, See Figure 7A. By combining a 3p19 antibody with an anti-TNFα antibody (e.g., 1:333 (w / w ) ratio), such a significant improvement in colon histopathology is This was unexpected. 500 μg / mouse TNFα + 1.5 μg / mouse anti-IL-23p1 Colon histopathological scores observed in the group receiving 9 were significantly higher than those observed in the isotype control group. It is also expected that the colon histopathology scores observed in the 2 groups were not statistically different. These results were consistent with a fixed high dose of TNFα mAb and a suboptimal low dose. Combination therapy with IL-23p19 significantly improved survival compared with monotherapy with either cytokine. have been shown to provide excellent protection.

[0161] Example 5: Combined anti-TNFα and anti-IL-23p19 therapy inhibits wound healing pathways influence on the unique subnetworks enriched by The molecular impact of combination therapy versus monotherapy with anti-TNFα and anti-IL-23p19 antibodies was determined. . Anti-TNFα (500μg) monotherapy or high-dose anti-IL-23p19 (25μg) monotherapy The humanized colonic gene expression signature of the method was compared with that of the combination therapy (500 μg anti-TNFα / 1.5 μg anti-TNFα). anti-TNFα antibody was crossed with the gene expression signature obtained from the anti-IL-23p19 in The molecular response to combination therapy with a low-dose anti-IL-23p19 antibody was compared with that of either treatment alone. The effect was determined to be additive or specific compared to the control group.

[0162] The 25 μg dose of anti-IL-23p19 treatment was selected for comparison to the suboptimal dose of anti-TNFα. The effect of combination therapy with anti-IL-23p19 at a dose that had efficacy in the model was assessed. The effect of monotherapy doses of anti-IL-23p19 was compared.

[0163] Similar to Study 1, we assessed signature overlap, generated treatment subnetworks, and analyzed enrichment solutions. To perform the analysis, humanized colon gene signatures were generated for each monotherapy and combination therapy treatment group. The data are shown in the left panel of Figure 8. TNFα or 25 μg of anti-IL-23p19 compared with combination therapy (500 μg of anti-T 220 genes were specifically and differentially expressed after NFα / 1.5 μg anti-IL-23p19 These genes were then analyzed using the CERTIFI gut Bayesian network. The maximum connected component of the resulting induced one-step subnetwork is defined as the richness The results are shown in the right panel of Figure 8. The analysis revealed that the combination treatment enriched in fibroblasts and extracellular matrix tissue. , specific sub-networks for cell types and pathways involved in wound repair and mucosal healing ( Therefore, anti-TNFα and anti-IL-23p19 therapy was identified. When used in combination, they may be able to identify both shared and unique disease-related pathways. Targeting the

[0164] Example 6: Clinical trial of anti-TNFα and anti-IL-23p19 therapy in UC Guselkumab and Golimumab in Participants With Moderately to Severely Active Ulcerative Colitis A randomized, phase 2a study to evaluate the efficacy and safety of combination therapy with rivaroxaban. A double-blind, active-controlled, parallel-group, multicenter, proof-of-concept clinical study Guselkumab (CNTO 1959 or TREMFYA®) is a human interferon-like compound. A complete antibody that binds with high specificity and affinity to the p19 subunit of leukin (IL)-23 It is a human immunoglobulin G1 lambda monoclonal antibody (mAb). Binding of IL-23 to cell surface IL-23 receptors promotes binding of extracellular IL-23 is blocked, inhibiting IL-23-specific intracellular signaling and subsequent cytokine production. Guselkumab is currently available in the United States, the European Union, Canada, and several other countries. Guselkumab is approved for the treatment of moderate to severe plaque psoriasis. It is also highly regarded worldwide for its use in the treatment of psoriatic arthritis (PsA) and Crohn's disease. .

[0165] Golimumab (CNTO 148 or SIMPONI®) has high TNFα activity. This is a fully human anti-tumor necrosis factor alpha (TNFα) mAb that binds with high affinity. Its action is to prevent the binding of TNFα to its receptor, thereby inhibiting the biological activity of TNFα. Golimumab is approved for the treatment of moderately to severely active ulcerative colitis in more than 90 countries worldwide. In addition, golimumab is approved for the treatment of rheumatoid arthritis (rheumatoid arthritis, RA), PsA, ankylosing spondylitis (AS), non-radiation nonradiographic axial spondyloarthritis (nr-Axial SpA, and polyarticular juvenile idiopathic arthritis It is approved in countries around the world for one or more of the following conditions: rheumatoid arthritis, rheumatoid arthritis, and pJIA.

[0166] Objectives and evaluation items The study consisted of two distinct phases: a 12-week combined comparison phase followed by a 26-week It consists of a week-long monotherapy phase.

[0167] the purpose Main purpose Combination Comparison Stage - Clinical efficacy of the combination therapy with guselkumab and golimumab was evaluated in patients with moderate to severe pulmonary embolism. To be evaluated in participants with active UC. The safety of combination therapy with guselkumab and golimumab was evaluated in patients with moderate to severe active To be evaluated in participants with UC.

[0168] secondary purpose Combination Comparison Stage To evaluate the effect of combination therapy with guselkumab and golimumab on endoscopic improvement do. Combination therapy with guselkumab and golimumab improves disease-specific health outcomes, including fatigue Evaluate the impact on health-related quality of life (HRQOL) do. The efficacy of the combination therapy with guselkumab and golimumab was evaluated in patients with negative baseline Assessed by sexual response signature status. C-reactive protein (CRP), fecal calprotectin, and Combination therapy with guselkumab and golimumab, including changes in other PD biomarkers Pharmacokinetics (PK), immunogenicity, and pharmacodynamics (PD) ) to evaluate.

[0169] Monotherapy phase To evaluate the clinical efficacy of the combination therapy followed by selcumab monotherapy. Evaluate the safety of the combination therapy followed by selcumab monotherapy. To evaluate the effect of combination therapy followed by selcumab monotherapy on endoscopic improvement. The effect of combination therapy followed by selcumab monotherapy on disease-specific HRQOL, including fatigue, was Evaluate the impact. Efficacy of the combination therapy followed by selkumaab monotherapy was assessed based on baseline negative response signals Evaluate based on sticky condition. Concomitant therapy, including changes in CRP, fecal calprotectin, and other PD biomarkers The objective of this study was to evaluate the PK, immunogenicity, and PD of the selkumaab monotherapy regimen followed by selkumaab monotherapy.

[0170] Experimental Purpose - Combination therapy is effective in patient-reported outcome (PRO) measures (e.g., , Bristol Stool Form Scale (BSFS) and severity of UC Patient's Global Impression of Change (PGIC) ) to investigate the impact on

[0171] Evaluation items Primary endpoint Clinical response at 12 weeks, measured by a difference from baseline in the Mayo score A reduction of 30% or more and 3 or more points in the rectal bleeding subscore (RBS) was A small number is defined as 1 or more, or an RBS of 0 or 1.

[0172] Key secondary endpoints Clinical remission at 12 weeks with a Mayo score of 2 or less and greater than 1 Defined as no individual subscore. Note: Other definitions of remission may be considered and should be included in the Statistical Analysis Plan. n, SAP).

[0173] hypothesis The combination of guselkumab and golimumab was superior to both monotherapy treatment arms12 The clinical response rate at week 1 is obtained.

[0174] Overall Plan This is a clinical trial of guselkumab and golimumab in adults with moderate to severe active UC. A Phase 2a, randomized, randomized, randomized controlled trial designed to evaluate the efficacy and safety of combination therapy with rivaroxaban. A randomized, double-blind, active-controlled, parallel-group, multicenter, interventional proof-of-concept (POC) study The target population is patients with moderate to severe active UC at baseline. Men or women aged 18-65 years with moderate to severe disease with a Mayo score of 6-12 and an endoscopic subscore of ≥2 obtained during central review of video endoscopy. Participants must be naïve to TNF antagonists and receive oral or intravenous Intravenous (IV) corticosteroids or immunomodulatory drugs (6-mercaptopurine have failed or become resistant to conventional treatment with 6-MP or azathioprine (AZA) do not have.

[0175] Immunomodulatory drugs (6-MP, AZA, and methotrexate (MTX)) were administered at the first dose. Must be discontinued for at least 2 weeks before study intervention. For participants receiving corticosteroids, the investigator will The daily dose of steroids must be tapered. All participants will be Clinical worsening of UC is assessed. Generally, the dose of combination therapy for UC is adjusted over a 38-week period. Patients should remain stable (oral corticosteroid tapering should begin at week 6) Concomitant treatment with UC is not deemed medically necessary by the investigator, except Initiation of prohibited therapy will result in discontinuation of the study intervention. do.

[0176] Centrally read endoscopies were performed at screening / baseline, week 12, and week 38. Consenting participants will have a further endoscopy at week 4, which will also be centrally read. Efficacy, PK and PD parameters, biomarkers, and safety will be assessed according to the Schedule of Activities (SoA). Blood samples for pharmacogenetics will be collected by consenting to this component of the protocol. Participation in pharmacogenetic research is optional. It is my intention.

[0177] Interim analyses are planned to inform future clinical developments. A database lock (DBL) is planned and all participants are safe. A final DBL is scheduled after completing the full follow-up visit. A Data Monitoring Committee (DMC) will be appointed.

[0178] Number of participants A target of 210 participants was enrolled in the study, with 70 participants planned per intervention group. It is being done.

[0179] Intervention group and period The study consisted of two distinct phases: a 12-week combination comparison phase followed by a 2-week The study consisted of a 6-week monotherapy phase. At week 0, 210 targeted participants were randomly assigned to receive Guselkumab. combination therapy with guselkumab and golimumab, guselkumab monotherapy, or golimumab monotherapy Randomized in a 1:1:1 ratio to either corticosteroids at baseline (Y / N). Participants randomized to combination therapy will be stratified by the use of steroids. Participants randomized to the monotherapy arm will then receive guselkumab monotherapy at week 12. The combination therapy arm will continue to receive the monotherapy initially randomized. To facilitate interpretation, the data for guselkumab and guselkumab used in each monotherapy intervention arm are The same dose regimen of limumab will be used. The following is a description of the three intervention groups. Combination therapy: guselkumab 200 mg IV and golimumab 200 mg subcutaneously at week 0 subcutaneous, SC); golimumab 100 mg SC at weeks 2, 6, and 10; and golimumab 100 mg SC at weeks 4 and 8. Guselkumab 200 mg IV every 8 weeks followed by guselkumab 100 mg IV every 8 weeks C; Guselkumab monotherapy: Guselkumab 200 mg IV at weeks 0, 4, and 8, followed by guselkumab 100 mg SC every 8 weeks; Golimumab monotherapy: Golimumab 200mg SC injection at week 0, followed by Golimumab 100 mg, then golimumab 100 mg every 4 weeks (q4w) In addition, placebo administration (IV or SS) was performed to maintain blinding throughout the study. C) is given as needed.

[0180] Overall participant duration is a maximum of 58 weeks in total (screening: maximum 8 weeks Treatment period: 38 weeks (12 weeks for the combination comparison phase and 26 weeks for the monotherapy phase) Safety follow-up: Approximately 16 weeks after the last dose of study intervention at Week 34. The end of the study will be This is defined as when a participant completes their last safety follow-up visit.

[0181] Efficacy evaluation (evaluation items) Efficacy assessment includes: Mayo score and partial Mayo score Ulcerative Colitis Endoscopic Index of Severity erity, UCEIS) Inflammatory PD markers including CRP and fecal calprotectin Patient-reported endpoints (i.e., patient-reported measures) to assess HRQOL outcomes and fatigue Inflammatory Bowel Disease Questionnaire (IBDQ), Patient-Reported Outcomes Measurement Information System stem, PROMIS-29, and PROMIS Fatigue 7-item Short Form (7a) Patient-reported symptom-exploratory scales, including BSFS and PGIC, for UC severity

[0182] Other efficacy evaluations (evaluation items) Efficacy assessment includes: Combination comparison phase (i.e., up to week 12) Endoscopic healing (endoscopic Mayo subscore 0 or 1) at 12 weeks. · Normalization of the endoscopic appearance of the mucosa (endoscopic Mayo subscore of 0). Histological healing at 12 weeks. Mucosal healing at 12 weeks (combined endoscopic and histological healing by Mayo) ). Inflammatory Bowel Disease Questionnaire (IBDQ) at 6 and 12 weeks Change from baseline in the Interventional Brain Development Questionnaire (IBDQ) total score. A >20-point improvement in IBDQ score at weeks 6 and 12. Patient-Reported Outcome Information System (PRI) data at 6 and 12 weeks PROMIS (Public Health Measurement Information System)-29 7 domains and abdominal Change from baseline in pain numerical rating scale. Fatigue response at 6 and 12 weeks (based on PROMIS Fatigue Short Form 7a; S (as defined in AP). Clinical response at week 12 according to baseline negative response signature status , clinical remission, and endoscopic cure. Change from baseline in Mayo score at week 12. Change from baseline in partial Mayo score through week 12. Change from baseline in CRP to week 12. Change from baseline in fecal calprotectin concentrations through week 12. CRP at week 12 in participants with abnormal CRP concentrations at baseline Concentration normalization. At week 12, participants with abnormal fecal calprotectin concentrations at baseline Normalization of fecal calprotectin concentrations at time points. At weeks 0 and 12 according to the level of Mayo endoscopy score at the corresponding visit Ulcerative Colitis Endoscopic Index of Severity (UCEIS) score at points. Change from baseline in UCEIS score at week 12. UCEIS score ≤ 4 at 12 weeks. UC-related emergency department visits, hospitalizations, and surgeries through week 12.

[0183] Monotherapy phase (i.e., week 12 and beyond) Clinical remission at 38 weeks. Clinical response at 38 weeks. Maintenance of clinical response at week 38 in participants who achieved a clinical response at week 12 Possessed. Endoscopic cure at 38 weeks. Normalization of the endoscopic appearance of the mucosa at 38 weeks. Histological healing at 38 weeks. Mucosal healing at 38 weeks. Clinical remission and no concomitant corticosteroids at 38 weeks. Maintenance of clinical remission at week 38 in participants who achieved clinical remission at week 12 Possessed. -Changes from baseline in IBDQ total score at 24 and 38 weeks change. >20-point improvement in IBDQ total score at 24 and 38 weeks good. PROMIS-29 7 domains and abdominal pain numerical assessment at 24 and 38 weeks Change from baseline in valence scales. Fatigue response at 24 and 38 weeks. Clinical response at week 38 according to baseline negative response signature status Answer: Clinical remission and endoscopic cure. Change from baseline in Mayo score at 38 weeks. Change from baseline in partial Mayo score through week 38. Change from baseline in CRP through week 38. Change from baseline in fecal calprotectin concentrations through week 38. CRP at week 38 in participants with abnormal CRP concentrations at baseline Concentration normalization. At week 38, participants with abnormal fecal calprotectin concentrations at baseline Normalization of fecal calprotectin concentrations at time points. U at 38 weeks according to the level of Mayo endoscopy score at 38 weeks CEIS score. Change from baseline in UCEIS score at week 38. UCEIS score ≤ 4 at 38 weeks. UC-related emergency department visits, hospitalizations, and surgeries through week 38.

[0184] Exploratory endpoints BSFS scores over time. · PGIC distribution of UC severity over time.

[0185] Pharmacokinetic and immunogenicity evaluation Serum samples were analyzed using a validated, specific, and sensitive immunoassay under sponsor supervision. As analyzed below, the concentrations of guselkumab and golimumab, and anti-guselkumab and anti-golimumab The detection of antibodies is assessed in each case.

[0186] Pharmacodynamic and biomarker evaluation Biomarker assessment examines the biological response to treatment and provides insight into the treatment of UC. This will be done to identify biomarkers associated with rucumab and / or golimumab. The titration involves the assessment of relevant biomarkers in serum, stool, whole blood, and mucosal biopsy samples (R NA (ribonucleic acid), histology, and single cell isolation).

[0187] Pharmacogenetic (DNA) evaluation Approximately 5 mL of pharmacogenomics total sample was used for genetic analysis as specified in the SoA. Blood samples will be collected. Only participants who signed a consent form to participate in the genetic evaluation will be given whole blood deoxyribonuclease (WBDE) samples. Ribonucleic acid (deoxyribonucleic acid, DNA) samples will be collected. Participation in the study is voluntary.

[0188] Safety evaluation Safety assessments conducted at each study visit included adverse events (AEs, and those occurring between assessment visits), tuberculosis (TB) evaluation and other infections Assessment of the patient, laboratory blood tests (hematology and chemistry), vital signs, assessment of suicidality surveillance of concomitant medications; observation of injection site reactions, AEs time-related to infusion, and / or hypersensitivity Includes sexual response.

[0189] statistical methods Determining sample size A sample size of 210 participants (70 per intervention group) was used to compare the combination therapy and both monotherapies. The significance of the proportion of participants in clinical response (primary endpoint) at 12 weeks between The power to detect differences was determined using a one-sided chi-square test with a significance level of 0.1 for each comparison. This study was designed to achieve both comparisons against monotherapy for the primary endpoint. Based on simulations, the combination therapy was sized to have approximately 80% power. The proportion of participants with clinical response at 12 weeks is expected to be 75% with combination therapy. This was based on the additive effect of both monotherapies (compared to a 35% historical placebo response). (20% improvement for each monotherapy).

[0190] Efficacy analysis All randomized participants receiving at least one study intervention were included in the efficacy analysis Participants will be included according to the treatment group to which they are randomized, regardless of the treatment they actually receive. Be analyzed.

[0191] For the primary endpoint of the study, the efficacy of the combination therapy will be compared to each monotherapy. For statistical comparison of both endpoints, concomitant use of corticosteroids at baseline (Y / N) was included. stratified by Cochran-Mantel-Haenszel (COH-Haenszel, The chi-square test (CMH) was used. Tests were performed simultaneously at a significance level of 0.1 for each comparison. This study demonstrated that the combination therapy group was significantly different from both monotherapy groups with respect to the primary endpoint. If the result is positive, it is considered positive.

[0192] If both tests for the primary endpoint were positive, corticosteroids were administered at baseline (Y / N). A one-sided CMH chi-square test stratified by id comorbidity was used to evaluate the primary secondary outcome. The efficacy of the combination therapy will be compared with each monotherapy on the endpoints. The study will be conducted at a significance level for each comparison. This is done simultaneously on either side of 0.1.

[0193] Analyses of other efficacy endpoints were performed without adjustment for multiple comparisons and were published. The nominal p-value is provided.

[0194] Safety analysis AE, serious adverse events (SAE), infection, serious infection, clinical Symptoms of flu-like symptoms include, but are not limited to, changes in laboratory evaluations and changes in vital signs. All data will be summarized. Treatment-emergent AEs will be divided by treatment group and by International Medical Glossary (Medical Dictionary for Regulatory Activities, MedDRA) system organ class and It is summarized by basic words.

[0195] Other analyses Pharmacokinetic analysis Serum guselkumab and golimumab concentrations over time were analyzed for each treatment using descriptive statistics. The data will be summarized over time for each treatment group.

[0196] Population PK modeling may be performed when appropriate. If applicable, the results of these analyses will be presented in a separate report.

[0197] Immunogenicity analysis The incidence of antibodies to guselkumab and golimumab was assessed by the Patients who have received at least one dose and have detectable antibodies to guselkumab and golimumab All participants with adequate samples for the study (i.e., one of guselkumab or golimumab) Participants with at least one sample obtained after the first administration will be summarized.

[0198] Pharmacokinetic / pharmacodynamic analysis Serum concentrations of guselkumab and golimumab, efficacy measurements and / or related biomarkers The relationships between the Analysis can be performed.

[0199] Biomarker analysis Serum protein samples, stool biomarkers, and biopsy and whole blood RNA obtained over time Changes in baseline and baseline values ​​for selected markers will be summarized by treatment group. The association between change from baseline and response to treatment will be investigated. -The analysis will be compiled in a separate technical report.

[0200] Pharmacogenetic analysis Genetic (DNA) analysis will be performed on participants who signed consent forms to participate in the pharmacogenomics substudy. These analyses are considered exploratory and will be compiled in a separate technical report. do.

[0201] This application describes several examples and embodiments of the present invention. and the like, without departing from the scope and essence of the present invention in principle. It should be noted that various modifications of the embodiment may be made. With this in mind, Other embodiments are within the scope of the items listed below. All numerical ranges given include all subranges contained therein as well as any subranges of those ranges. All publications and patents mentioned herein include any individual value within a range. The application is incorporated herein by reference.

[0202] The present invention can be described with reference to the following numbered embodiments. 1. IL-23 inhibitors and TN inhibitors for use in treating inflammatory diseases in patients Use of an Fα inhibitor, wherein the inhibitor is in a co-therapeutically effective amount and the patient shows a clinical response. IL-23 inhibitors and TNFα inhibitors for 2. The inflammatory disease is inflammatory bowel disease, and the patient has a Mayo score, partial Mayo score, or A. Ulcerative Colitis Endoscopic Severity Index (UCEIS), marker CRP and / or fecal Calprotectin and clinical trials selected from the group consisting of patient-reported outcome and symptom scales IL-23 inhibition for use according to embodiment 1, which shows a clinical response based on the endpoint agents and TNFα inhibitors. 3. The IL-23 inhibitor comprises an anti-IL-23p19 antibody or an antigen-binding fragment thereof, and 3. The method of claim 1 or 2, wherein the Fα inhibitor comprises an anti-TNFα antibody or an antigen-binding fragment thereof. An IL-23 inhibitor and a TNFα inhibitor for use. 4. The method of any one of embodiments 1 to 3, wherein the inflammatory bowel disease is Crohn's disease. IL-23 inhibitor and TNFα inhibitor for 5. Embodiments 1-4, wherein the inflammatory bowel disease is ulcerative colitis (UC) or unclassifiable colitis. 1. An IL-23 inhibitor and a TNFα inhibitor for use according to any one of the above. Embodiment 6. The inflammatory bowel disease is moderate to severe active ulcerative colitis (UC). The use of an IL-23 inhibitor and a TNFα inhibitor according to any one of embodiments 1 to 5. Harmful agent. 7. Patient has been previously treated with a TNFα inhibitor alone and UC is in remission after the previous treatment. IL-23 inhibitor for use according to any one of embodiments 1 to 6, and TNFα inhibitors. 8. Patient has been previously treated with an IL-23 inhibitor alone and UC has not progressed beyond the previous treatment IL-23 inhibition for use according to any one of embodiments 1 to 7, which did not result in remission agents and TNFα inhibitors. 9. The anti-IL-23p19 antibody comprises: a) a heavy chain complementarity-determining region (CDR) of SEQ ID NOs: 1 to 3 a) the amino acid sequence and the light chain CDR amino acid sequence of SEQ ID NO: 4 to 6, b) the heavy chain variable a) a heavy chain variable region amino acid sequence of SEQ ID NO: 9 and a light chain variable region amino acid sequence of SEQ ID NO: 10; 9. Any of embodiments 1 to 8, comprising a light chain amino acid sequence of SEQ ID NO: 10 and a light chain amino acid sequence of SEQ ID NO: 11. 1. An IL-23 inhibitor and a TNFα inhibitor for use as described in claim 1. 10. An anti-TNFα antibody comprising: a) a heavy chain CDR amino acid sequence of SEQ ID NOs: 11 to 13 and a sequence a) light chain CDR amino acid sequences of SEQ ID NO: 14 to 16, b) heavy chain variable region amino acid sequence of SEQ ID NO: 17 and the light chain variable region amino acid sequence of SEQ ID NO: 18, or c) the heavy chain amino acid sequence of SEQ ID NO: 19. 10. The method of claim 1, further comprising administering to said patient a human immunodeficiency virus (HIV) infection, comprising administering to said patient a human immunodeficiency virus (HIV) infection, and a human ... IL-23 inhibitor and TNFα inhibitor for use in 11. The anti-IL-23p19 antibody comprises: a) a heavy chain complementarity determining region (CDR) of SEQ ID NOs: 1 to 3; a) the amino acid sequence and the light chain CDR amino acid sequence of SEQ ID NO: 4 to 6, b) the heavy chain CDR amino acid sequence of SEQ ID NO: 7 the light chain variable region amino acid sequence of SEQ ID NO: 8, or c) the light chain variable region amino acid sequence of SEQ ID NO: 9 the heavy chain amino acid sequence of SEQ ID NO: 10 and the light chain amino acid sequence of SEQ ID NO: 11, The heavy chain CDR amino acid sequences of SEQ ID NOS: 11 to 13 and the light chain CDR amino acid sequences of SEQ ID NOS: 14 to 16 a) the heavy chain variable region amino acid sequence of SEQ ID NO: 17 and the light chain variable region amino acid sequence of SEQ ID NO: 18 or c) a heavy chain amino acid sequence of SEQ ID NO: 19 and a light chain amino acid sequence of SEQ ID NO: 20 11. The method for inhibiting IL-23 for use according to any one of embodiments 1 to 10, comprising the sequence agents and TNFα inhibitors. 12. An anti-IL-23 antibody or antibody for use in the treatment of ulcerative colitis in a patient and an anti-TNFα antibody or a fragment thereof, wherein the anti-IL-23p19 antibody is (i) Heavy chain complementarity determining region (CDR) amino acid sequences of SEQ ID NOS: 1 to 3 and light chains of SEQ ID NOS: 4 to 6 CDR amino acid sequence, (ii) the heavy chain variable region amino acid sequence of SEQ ID NO: 7 and SEQ ID NO: 8 a light chain variable region amino acid sequence of SEQ ID NO: 9, or (iii) a heavy chain amino acid sequence of SEQ ID NO: 9 10 light chain amino acid sequences, and the anti-TNFα antibody comprises (i) a heavy chain of SEQ ID NOs: 11 to 13 CDR amino acid sequences and light chain CDR amino acid sequences of SEQ ID NOs: 14 to 16, (ii) SEQ ID NOs: the heavy chain variable region amino acid sequence of SEQ ID NO: 17 and the light chain variable region amino acid sequence of SEQ ID NO: 18; or (iii) a heavy chain amino acid sequence of SEQ ID NO: 19 and a light chain amino acid sequence of SEQ ID NO: 20. and the antibody is in a co-therapeutically effective amount, and the use is effective in treating ulcerative colitis, and the patient The subjects were assessed by Mayo score, partial Mayo score, and endoscopic severity index of ulcerative colitis (U CEIS), markers CRP and / or fecal calprotectin, and patient-reported outcomes and a symptom scale, and an anti-IL-23 antibody or a fragment thereof and an anti-TNFα antibody or a fragment thereof for the treatment of IL-23. 13. Anti-TNFα antibody and anti-IL-23p19 antibody are administered at a ratio of 1:2 to 2:1 (w / w). 13. The method of claim 12, wherein the anti-IL-23 antibody and the anti-TNFα antibody are administered intravenously. body. 14. The anti-TNFα antibody and the anti-IL-23p19 antibody are mixed in a ratio of 15:1 to 400:1 (w / w 14. The anti-IL-23 antibody and method for use according to embodiment 12 or 13, wherein the antibody is administered in a ratio of 0.5:1 (IL-23 / IL-23) / ( ... and anti-TNFα antibody. 15. Embodiment 1, in which the anti-IL-23p19 antibody and the anti-TNFα antibody are administered simultaneously. 15. An anti-IL-23 antibody and an anti-TNFα antibody for use according to any one of 2 to 14. 16. Embodiment 12, in which the anti-IL-23p19 antibody and the anti-TNFα antibody are administered sequentially. 15. An anti-IL-23 antibody and an anti-TNFα antibody for use according to any one of claims 1 to 14. 17. The anti-IL-23p19 antibody and the anti-TNFα antibody are administered within one day of each other. The anti-IL-23 antibody and and anti-TNFα antibody. 18.Anti-IL-23p19 antibody is administered at an initial intravenous dose of 200 mg, at weeks 4 and 8. administered as an intravenous dose of 200 mg every 8 weeks, followed by subcutaneous doses of 100 mg every 8 weeks. The anti-TNFα antibody was administered at an initial subcutaneous dose of 200 mg and at 100 mg at weeks 2, 6, and 10. 18. The method of claim 12, wherein the medicament is administered in a subcutaneous dose of 1 mg. Anti-IL-23 antibody and anti-TNFα antibody for the treatment of rheumatoid arthritis. 19. The patient has a Mayo score, partial Mayo score, or endoscopic severity of ulcerative colitis. The UCEIS, markers CRP and / or fecal calprotectin, and patient reports Clinical remission was assessed based on clinical endpoints selected from a group consisting of clinical outcomes and symptom scales. 19. An anti-IL-23 antibody for use according to any one of embodiments 12 to 18, Anti-TNFα antibody. 20. The method of embodiment 19, wherein the clinical endpoint is measured about 12 weeks after initial treatment. An anti-IL-23 antibody and an anti-TNFα antibody for use. 21. The method of embodiment 19 or 20, wherein the clinical endpoint is based on the Mayo score. An anti-IL-23 antibody and an anti-TNFα antibody for use. 22. Antibiotics for use in reducing colonic inflammation in patients with inflammatory bowel disease an IL-23 antibody or a fragment thereof and an anti-TNFα antibody or a fragment thereof, The amount is effective, and the use reduces inflammation in the patient's colon to a level comparable to that of the colon of a normal subject. and an anti-IL-23 antibody or fragment thereof and an anti-TNFα antibody or fragment thereof, which are effective for is a fragment of it. 23. Anti-IL-23p19 antibody or its antigen-binding fragment and anti-TNFα antibody or its antigen After administration of the binding fragment, inflammation is minimal or normal in tissue samples from the patient's colon. 23. An anti-IL-23 antibody and an anti-TNFα antibody for use according to embodiment 22. 24. Anti-IL-23p19 antibody or its antigen-binding fragment and anti-TNFα antibody or its antigen Following administration of the binding fragment, glandular loss is minimal or normal in tissue samples from the colon of the subject. 23. An anti-IL-23 antibody and an anti-TNFα antibody for use according to embodiment 22. 25. Anti-IL-23p19 antibody or its antigen-binding fragment and anti-TNFα antibody or its antigen After administration of the binding fragment, erosion is minimal or normal in tissue samples from the colon of the subject. 23. An anti-IL-23 antibody and an anti-TNFα antibody for use according to embodiment 22. 26. Anti-IL-23p19 antibody or its antigen-binding fragment and anti-TNFα antibody or its antigen Following administration of the binding fragment, mucosal thickness and hyperplasia were independently measured in tissue samples from the colon of the subjects. 23. The anti-IL-23 antibody and anti-IL-23 antibody for use according to embodiment 22, TNFα antibody. 27. Anti-IL-23p19 antibody or its antigen-binding fragment and anti-TNFα antibody or its antigen

[0033] Embodiment 22, wherein after administration of the binding fragment, the histopathology of the colon is identical to the histopathology of normal tissue. An anti-IL-23 antibody and an anti-TNFα antibody for use as described in 1. 28. The anti-IL-23p19 antibody or antigen-binding fragment thereof comprises: a) a heavy chain C of SEQ ID NOs: 1 to 3 a) a DR amino acid sequence and a light chain CDR amino acid sequence of SEQ ID NO: 4 to 6, b) a heavy chain CDR amino acid sequence of SEQ ID NO: 7 the variable region amino acid sequence of SEQ ID NO: 8 and the light chain variable region amino acid sequence of SEQ ID NO: 9; and a light chain amino acid sequence of SEQ ID NO: 10. d) the heavy chain CDR amino acid sequences of SEQ ID NOs: 11 to 13 and SEQ ID NO: 14 a) the light chain CDR amino acid sequence of SEQ ID NO: 16; b) the heavy chain variable region amino acid sequence of SEQ ID NO: 17; f) the light chain variable region amino acid sequence of SEQ ID NO: 18, or f) the heavy chain amino acid sequence of SEQ ID NO: 19, and The use according to any one of embodiments 22 to 27, comprising a light chain amino acid sequence of SEQ ID NO: 20. Anti-IL-23 antibody and anti-TNFα antibody for use in the treatment of rheumatoid arthritis. 29. Anti-TNFα antibody or its antigen-binding fragment and anti-IL-23p19 antibody or its antigen 29. Any of embodiments 22-28, wherein the binding fragments are administered in a ratio of 1:2 to 2:1 (w / w). An anti-IL-23 antibody and an anti-TNFα antibody for use according to any one of the preceding claims. 30. Anti-TNFα antibody or its antigen-binding fragment and anti-IL-23p19 antibody or its antigen 29. The method of claim 22, wherein the binding fragments are administered in a ratio of 15:1 to 400:1 (w / w). An anti-IL-23 antibody and an anti-TNFα antibody for use according to any one of the above. 31. a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof, and b) an anti-TNFα antibody or and the antigen-binding fragments thereof are administered simultaneously. An anti-IL-23 antibody and an anti-TNFα antibody for use. 32. a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof and b) an anti-TNFα antibody or The method of any one of embodiments 22 to 30, wherein the antigen-binding fragments are administered sequentially. Anti-IL-23 antibody and anti-TNFα antibody for the treatment of rheumatoid arthritis. 33. a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof and b) an anti-TNFα antibody or 31. The method of any one of embodiments 22 to 30, wherein the antigen-binding fragments are administered within one day of each other. An anti-IL-23 antibody and an anti-TNFα antibody for use as described. 34. For use in treating inflammatory bowel disease in a patient and reducing weight loss in a patient. and an anti-IL-23 antibody or a fragment thereof and an anti-TNFα antibody or a fragment thereof. 35. Anti-TNFα antibody or its antigen-binding fragment and anti-IL-23p19 antibody or its antigen 35. The method of claim 34, wherein the binding fragments are administered in a ratio of 15:1 to 400:1 (w / w). Anti-IL-23 antibody or antigen-binding fragment thereof and anti-TNFα antibody or antigen-binding fragment thereof for use Combined fragments. 36. a) anti-IL-23p19 antibody or its antigen-binding fragment and b) anti-TNFα antibody or 36. The method of claim 34, wherein the antigen-binding fragments thereof are administered simultaneously. An anti-IL-23 antibody or an antigen-binding fragment thereof and an anti-TNFα antibody or an antigen-binding fragment thereof. 37. a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof and b) an anti-TNFα antibody or 36. The antibody for use according to embodiment 34 or 35, wherein the antigen-binding fragments thereof are administered sequentially. An IL-23 antibody or an antigen-binding fragment thereof and an anti-TNFα antibody or an antigen-binding fragment thereof. 38. a) an anti-IL-23p19 antibody or an antigen-binding fragment thereof and b) an anti-TNFα antibody or 38. Any of embodiments 34, 35, or 37, wherein the antigen-binding fragments are administered within one day of each other. an anti-IL-23 antibody or an antigen-binding fragment thereof and an anti-TNFα antibody for use according to any one of claims 1 to 4; An antibody or antigen-binding fragment thereof. 39. The anti-IL-23p19 antibody or antigen-binding fragment thereof comprises: a) a heavy chain C of SEQ ID NOs: 1 to 3 a) a DR amino acid sequence and a light chain CDR amino acid sequence of SEQ ID NO: 4 to 6, b) a heavy chain CDR amino acid sequence of SEQ ID NO: 7 the variable region amino acid sequence of SEQ ID NO: 8 and the light chain variable region amino acid sequence of SEQ ID NO: 9; and a light chain amino acid sequence of SEQ ID NO: 10, d) the heavy chain CDR amino acid sequences of SEQ ID NOs: 11 to 13 and SEQ ID NO: 1 e) a light chain CDR amino acid sequence of SEQ ID NO: 4 to 16, f) a heavy chain variable region amino acid sequence of SEQ ID NO: 17, and f) the light chain variable region amino acid sequence of SEQ ID NO: 18, or f) the heavy chain amino acid sequence of SEQ ID NO: 19, and a light chain amino acid sequence of SEQ ID NO: 20. Anti-IL-23 antibody or antigen-binding fragment thereof and anti-TNFα antibody or antigen-binding fragment thereof for use in Original combined fragment. 40. For use in treating moderately to severely active ulcerative colitis in human patients an anti-IL-23 antibody or a fragment thereof and an anti-TNFα antibody or a fragment thereof for -23p19 antibody or its antigen-binding fragment is administered at 0.0005 to 0.002 mg / kg and (i) a heavy chain CDR amino acid sequence of SEQ ID NOs: 1 to 3 and a light chain CDR amino acid sequence of SEQ ID NOs: 4 to 6. CDR amino acid sequence, (ii) the heavy chain variable region amino acid sequence of SEQ ID NO: 7 and SEQ ID NO: 8 a light chain variable region amino acid sequence of SEQ ID NO: 9, or (iii) a heavy chain amino acid sequence of SEQ ID NO: 9 10 light chain amino acid sequences, and the anti-TNFα antibody or antigen-binding fragment thereof is 20 to 0.125 mg / kg, and (iv) a heavy chain CD of SEQ ID NOs: 11 to 13. (v) the light chain CDR amino acid sequence of SEQ ID NO: 17; and the heavy chain variable region amino acid sequence of SEQ ID NO: 18, or 19) a heavy chain amino acid sequence and SEQ ID NO: 20) a light chain amino acid sequence; An anti-IL-23 antibody or a fragment thereof and an anti-TNFα antibody or a fragment thereof for use. 41. The use according to embodiment 40, wherein the use is effective for treating ulcerative colitis. an anti-IL-23 antibody or an antigen-binding fragment thereof and an anti-TNFα antibody or an antigen-binding fragment thereof for use in Piece. 42. The patient has a Mayo score, partial Mayo score, or endoscopic severity of ulcerative colitis. The UCEIS, markers CRP and / or fecal calprotectin, and patient reports Clinical remission was assessed based on clinical endpoints selected from a group consisting of clinical outcomes and symptom scales. 42. The anti-IL-23 antibody or antigen binding thereof for use according to embodiment 40 or 41. and an anti-TNFα antibody or antigen-binding fragment thereof. 43. The anti-IL-23p19 antibody is a pharmaceutical composition containing 7.9% (w / v) sucrose, 4 0.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate; 0.053% ( Polysorbate 80 (w / v) present at 100 mg / mL in an aqueous solution of a composition containing The TNFα antibody was added to the pharmaceutical composition in a 4.1% (w / v) sorbitol, 5.6 mM L-hydroxybenzoate, and 1% (w / v) L-hydroxybenzoate. L-histidine and L-histidine monohydrochloride monohydrate; 0.015% (w / v) Polysorbate 80, wherein the composition is present at 100 mg / mL in an aqueous solution. 1. The anti-IL-23 antibody or antigen-binding fragment thereof for use according to any one of claims 1 to 2 and an anti-IL-23 antibody or antigen-binding fragment thereof for use according to any one of claims 1 to 2. TNFα antibody or antigen-binding fragment thereof.

Claims

[Claim 1] The invention described in the specification.

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