Α4β7 inhibitor and il-23 inhibitor combination therapy

Abstract

To provide a method for treating an autoimmune disorder and / or an inflammatory bowel disease.SOLUTION: Provided is a combination therapy comprising an α4β7 inhibitor, for example, an anti-α4β7 antibody, for example, vedolizumab, and an IL-23 inhibitor, for example, an anti-IL-23 antibody.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application is a continuation of U.S. Provisional Patent Application No. 62 / 835,349, filed April 17, 2019. No. 60 / 019,493, filed on May 1, 2006, the contents of which are incorporated herein by reference.

[0002] Sequence Listing This application contains herewith a Sequence Listing which is submitted in electronically readable format. The sequence listing file was created on April 14, 2020, and is named "T103022_1100WO The file is named "_0415_4_SL.txt" and is 24kb in size. The entire contents of the sequence listing in sequencelisting.txt are incorporated herein by reference. It can be enjoyed.

[0003] Technical Field The present invention relates to α4β7 inhibitors, such as anti-α4β7 antibodies (e.g., vedolizumab), and Methods and compositions relating to combination therapies including IL-23 inhibitors, e.g., anti-IL-23 antibodies Pertaining to things. [Background technology]

[0004] Interleukin-23 (IL-23) is composed of the p40 and p19 subunits. IL-23 is a heterodimeric cytokine composed of IL-23 and IL-25. and in response to infection with fungal pathogens, antigen-presenting cells (e.g., dendritic cells and macrophages) IL-23R, the IL-23 receptor, is produced by Th17 cells and monocytes. cells, γδT cells, natural killer (NK) cells, dendritic cells, macrophages, and autologous It is expressed on a variety of adaptive and innate immune cells, including natural lymphocytes. Effector cytokines play a key role in the pathogenesis of inflammatory bowel disease (IBD) in acute and chronic mouse models. D) plays an important role in the pathogenesis of IBD. In IBD patients, IL-23R gene expression and tandem Protein levels are elevated at the intestinal mucosal surface.

[0005] IL-23 inhibitors represent an exciting new class of targeted molecules for treating IBD For example, monoclonal antibodies against IL-23 inhibit the T cell proliferation of naive T cells. H To 17 cells It has been shown to limit differentiation of IBD cells, thereby ameliorating the pathogenesis of IBD (e.g., W See O2018 / 112232).

[0006] Targeted IL-23 therapy (Kashani and Schwartz (2019)Gas troenterol Hepatol NY 15(5):255 Regardless of the cause, it can reduce the disease burden, improve remission rates, and slow progression of immune disorders such as IBD. There remains an unmet medical need for alternative therapies to achieve this. Summary of the Invention

[0007] In various aspects, the present disclosure provides an α4β7 inhibitor, such as an anti-α4β7 antibody, or an antigen-binding IL-23 inhibitors, such as anti-IL-23 antibodies or their antigen-binding fragments The present invention relates to the treatment of autoimmune disorders and / or inflammatory bowel disease by administering a combination of Provide a method for placing

[0008] In one aspect of the invention, provided herein is a method of treating a human patient in need thereof. The method includes administering to the human patient an α4β7 inhibitor and an IL-23 inhibitor.

[0009] In one embodiment, the α4β7 inhibitor is an anti-α4β7 antibody. The β7 antibody is humanized. In one embodiment, the anti-α4β7 antibody is C4β7 as set forth in SEQ ID NO: 4. DR3 domain, CDR2 domain set forth in SEQ ID NO: 3, and CDR set forth in SEQ ID NO: 2 a heavy chain variable region comprising a CDR3 domain set forth in SEQ ID NO: 8; a CDR4 domain set forth in SEQ ID NO: 9; 7, and a light chain variable region comprising a CDR2 domain as set forth in SEQ ID NO: 6. Includes the area.

[0010] In one aspect of the invention, provided herein is a method of treating a human patient in need thereof. The method comprises administering to a human patient an anti-α4β7 antibody and an IL-23 inhibitor, The α4β7 antibody is an IgG1 antibody; the CDR3 domain is set forth in SEQ ID NO: 4, SEQ ID NO: 3 and a CDR1 domain as set forth in SEQ ID NO: 2. the CDR3 domain set forth in SEQ ID NO: 8, the CDR2 domain set forth in SEQ ID NO: 7 and a light chain variable region comprising the CDR1 domain set forth in SEQ ID NO:6.

[0011] In one embodiment, the human patient has an autoimmune disease. In certain embodiments, the autoimmune disease is rheumatoid arthritis, psoriasis, or arthritis. psoriatic arthritis, or axial spondyloarthritis.

[0012] In one embodiment, the human patient has inflammatory bowel disease (IBD). BD is a condition characterized by ulcerative colitis (e.g., moderately to severely active ulcerative colitis) or Crohn's disease. (e.g., moderate to severe active Crohn's disease).

[0013] In one embodiment, the anti-α4β7 antibody is administered before the IL-23 inhibitor.

[0014] In another embodiment, the anti-α4β7 antibody is administered after the IL-23 inhibitor.

[0015] In one embodiment, the anti-α4β7 antibody is administered simultaneously with the IL-23 inhibitor.

[0016] In another embodiment, the anti-α4β7 antibody has a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:1. The present invention relates to a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:5.

[0017] In one embodiment, the anti-α4β7 antibody is a humanized antibody.

[0018] In one embodiment, the anti-α4β7 antibody is vedolizumab.

[0019] In one embodiment, a human patient receives an initial dose of 300 mg of anti-α4β7 antibody at week 0, followed by , a second dose of 300 mg of anti-α4β7 antibody was administered at week 2, followed by a second dose of 300 mg of anti-α4β7 antibody at week 6. In one embodiment, the human patient is administered a third dose of 00 mg. Thereafter, 300 mg of anti-α4β7 antibody is administered every 8 weeks. Patients will receive 300 mg of anti-α4β7 antibody every 4 weeks if they do not show clinical improvement. .

[0020] In another embodiment, the compound is administered to a human patient, the human patient having ulcerative colitis or Crohn's disease. and clinical improvement is clinical remission.

[0021] In one embodiment, the human patient receives an anti-α4β7 antibody every four weeks starting eight weeks after the third dose. The patient is given 300mg.

[0022] In an alternative embodiment, the human patient receives anti-α4β7 every two weeks starting eight weeks after the third dose. The patient was given 108 mg of antibody.

[0023] In one embodiment, a human patient receives an initial dose of 300 mg of anti-α4β7 antibody at week 0, followed by , a second dose of 300 mg of anti-α4β7 antibody in the second week, followed by 1 dose of anti-α4β7 antibody in the sixth week. A third dose of 0.08 mg will be administered, followed by a dose of 108 mg every two weeks thereafter.

[0024] In yet another embodiment, the anti-α4β7 antibody is administered intravenously to a human patient. In an embodiment, a 300 mg dose is administered intravenously.

[0025] In one embodiment, the anti-α4β7 antibody is administered subcutaneously to a human patient. A dose of 108 mg is administered subcutaneously.

[0026] In one embodiment, the IL-23 inhibitor is an antibody that binds to the p19 subunit of IL-23. It is the body.

[0027] In one embodiment, the IL-23 inhibitor is an antibody that binds to the p40 subunit of IL-23. It is the body.

[0028] In one embodiment, the IL-23 inhibitor binds to the p19 and p40 subunits of IL-23. It is an antibody that binds to the antibody.

[0029] In certain embodiments, the IL-23 inhibitor includes, but is not limited to, risankizumab, uste An anti-IL-23 antibody, such as kinumab, guselkumab, or tildrakizumab.

[0030] In one embodiment, the IL-23 inhibitor is an antibody that binds to IL-23R.

[0031] In one embodiment, the human patient is treated with an α4β7 inhibitor (e.g., an anti-α4β7 antibody, e.g., Non-responders or non-remissions at 6 and / or 10 weeks after initiation of treatment with dolizumab is characterized as a solver.

[0032] In one embodiment, the human patient is treated with an α4β7 inhibitor (e.g., an anti-α4β7 antibody, e.g., Serum I levels at the start of treatment with dolizumab or at 6 or 10 weeks after the start of treatment It has been characterized as having elevated levels of L-22.

[0033] In one embodiment, IL-22 levels in the serum of a human patient are measured by the administration of an anti-α4β7 antibody (e.g., From the start of treatment with vedolizumab to 10 weeks after the start, and from the start of treatment with anti-α4β7 antibody At the start of treatment and 6 weeks after the start, or 6 to 10 weeks after the start of treatment with anti-α4β7 antibody , not reduced at all, reduced by less than half, or reduced by less than one-third.

[0034] In a further embodiment, the human patient is treated with an anti-α4β7 antibody (e.g., vedolizumab). This is characterized by elevated serum levels of IL-22 at the start of treatment or six weeks after initiation of treatment. The serum IL-22 levels were measured at the start of treatment and at week 10, week 6, and week 7, respectively. or between weeks 6 and 10, there is no reduction, a reduction of less than half, or a reduction of a third. decreases to less than 1.

[0035] In one embodiment, the human patient is treated with an α4β7 inhibitor (e.g., an anti-α4β7 antibody, e.g., Serum I levels at the start of treatment with dolizumab or at 6 or 10 weeks after the start of treatment It has been characterized as having elevated levels of L-1β.

[0036] In a further embodiment, the human patient is treated with an anti-α4β7 antibody (e.g., vedolizumab). Elevated fecal calprotectin levels at the start of treatment or 6 weeks after initiation The fecal calprotectin levels were characterized as No reduction or less than half reduction between 6 weeks and 10 weeks, or reduced by less than one-third.

[0037] In certain embodiments, the human patient has elevated levels of IL-22, STAT5A, or IL-22 compared to control levels. In certain embodiments, the patient is characterized by elevated levels of IL-1β and / or IL-1β. , the human patient is treated with IL-22, STAT5A, and / or IL-12 as compared to a reference level. It is characterized by elevated levels of 1β.

[0038] In one embodiment, the control levels of IL-22, STAT5A, and / or IL-1β are , subjects not affected by IBD, healthy subjects, non-inflamed colon tissue from patients, or non-colon tissue. In one embodiment, the level of any one or more of IL-22 and / or The reference level of STAT5A is the starting level in patients who respond to treatment with vedolizumab. The level is determined by

[0039] In one embodiment, the patient's IL-22, STAT5A, and / or IL-1β levels are , measured initially, for example, before treatment with vedolizumab or on the first day of treatment.

[0040] In another embodiment, the patient's IL-22, STAT5A, and / or IL-1β levels For example, the starting level is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, Measured 8, 9, and / or 10 days prior.

[0041] In one embodiment, the patient's IL-22, STAT5A, and / or IL-1β nucleic acids and In certain embodiments, IL-22, STAT, and / or protein levels are measured. 5A and / or IL-1β levels are 5% higher than control or reference levels , 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80% In certain embodiments, the levels of IL-22, ST are increased by 90%, 100%, or more. AT5A and / or IL-1β levels compared to control or reference levels 5%~35%, 5%~25%, 5%~20%, 5%~15%, 5%~10%, 10%~2 0%, 10%-15%, 10%-30%, 15%-40%, 20%-50%, 25%-6 It can increase by 0%, 30%-70%, or 40%-100% or more.

[0042] In some aspects, the present invention provides a method of treating inflammatory bowel disease in a patient in need thereof. The method includes combining an anti-α4β7 antibody (e.g., vedolizumab) and a p19 subunit of IL-23. and administering to the patient an antibody that binds to the subunit, wherein the anti-α4β7 antibody is administered to the human patient. The first dose of anti-α4β7 antibody was 300 mg at week 0, followed by 30 mg of anti-α4β7 antibody at week 2. The second dose of 0 mg was administered at week 6, followed by a third dose of 300 mg of anti-α4β7 antibody at week 6. Starting 8 weeks after administration, patients were administered 300 mg of anti-α4β7 antibody every 8 weeks, and anti-α4β Antibody No. 7 has a CDR3 domain set forth in SEQ ID NO: 4 and a CDR2 domain set forth in SEQ ID NO: 3. and a heavy chain variable region comprising the CDR1 domain set forth in SEQ ID NO: 2; the CDR3 domain described above, the CDR2 domain described above in SEQ ID NO: 7, and the CDR3 domain described above in SEQ ID NO: 6 and a light chain variable region comprising the CDR1 domain, and wherein the patient has an increased level of IL-2 compared to a control level. 2, and is characterized by elevated levels of STAT5A and / or IL-1β.

[0043] In some aspects, the present invention provides a method of treating inflammatory bowel disease in a patient in need thereof. The method includes combining an anti-α4β7 antibody (e.g., vedolizumab) and a p40 subunit of IL-23. and administering to the patient an antibody that binds to the subunit, wherein the anti-α4β7 antibody is administered to the human patient. The first dose of anti-α4β7 antibody was 300 mg at week 0, followed by 30 mg of anti-α4β7 antibody at week 2. The second dose of 0 mg was administered at week 6, followed by a third dose of 300 mg of anti-α4β7 antibody at week 6. Starting 8 weeks after administration, patients were administered 300 mg of anti-α4β7 antibody every 8 weeks, and anti-α4β Antibody No. 7 has a CDR3 domain set forth in SEQ ID NO: 4 and a CDR2 domain set forth in SEQ ID NO: 3. and a heavy chain variable region comprising the CDR1 domain set forth in SEQ ID NO: 2; the CDR3 domain described above, the CDR2 domain described above in SEQ ID NO: 7, and the CDR3 domain described above in SEQ ID NO: 6 and a light chain variable region comprising the CDR1 domain, and wherein the patient has an increased level of IL-2 compared to a control level. 2, and is characterized by elevated levels of STAT5A and / or IL-1β.

[0044] In some aspects, the present invention provides a method of treating inflammatory bowel disease in a patient in need thereof. The method includes combining an anti-α4β7 antibody (e.g., vedolizumab) and a p19 subunit of IL-23. the patient is administered an antibody that binds to the subunit, and the anti-α4β7 antibody binds to the subunit at week 0. First dose of 300 mg of α4β7 antibody, followed by two doses of 300 mg of anti-α4β7 antibody in the second week 1 dose, followed by a third dose of 108 mg of anti-α4β7 antibody at week 6, followed by a second dose at week 2. The anti-α4β7 antibody contained the CDR3 domain set forth in SEQ ID NO: 4. a CDR1 domain as set forth in SEQ ID NO:2; a CDR2 domain as set forth in SEQ ID NO:3; and a CDR1 domain as set forth in SEQ ID NO:2. a heavy chain variable region comprising the CDR3 domain set forth in SEQ ID NO: 8, the CDR4 domain set forth in SEQ ID NO: 7; and a light chain variable region comprising the CDR2 domain of SEQ ID NO: 6 and the CDR1 domain of SEQ ID NO: 7. Patients had elevated levels of IL-22, STAT5A, and / or IL-1 compared to control levels. It is characterized by elevated levels of β.

[0045] In some aspects, the present invention provides a method for treating inflammatory bowel disease in a patient in need of such treatment. The present invention provides a method of treating a tumor in the lungs, the method comprising administering an anti-α4β7 antibody (e.g., vedolizumab) and IL- administering to a patient an antibody that binds to the p40 subunit of α4β7; The initial administration of anti-α4β7 antibody 300 mg was given at week 0, followed by anti-α4β7 antibody 3 mg at week 2. A second dose of 100 mg was administered at week 6, followed by a third dose of 108 mg of anti-α4β7 antibody at week 6, followed by and administered at a dose of 108 mg every two weeks thereafter. The anti-α4β7 antibody was the CDR3 domain described above, the CDR2 domain described above in SEQ ID NO: 3, and the CDR3 domain described above in SEQ ID NO: 2 a heavy chain variable region comprising a CDR1 domain; a CDR3 domain set forth in SEQ ID NO: 8; A light antibody comprising the CDR2 domain set forth in column number 7 and the CDR1 domain set forth in SEQ ID NO: 6. and the patient has elevated levels of IL-22, STAT5A, and / or IL-12, compared to control levels. or characterized by elevated levels of IL-1β.

[0046] In certain embodiments, the control level is determined from a subject not suffering from IBD, a healthy subject, a patient-derived subject, or a control group. The present invention relates to the treatment of colonic inflammation, which may be at one or more levels of normal, non-inflamed colonic tissue, or non-colonic tissue. [Brief explanation of the drawings]

[0047] [Figure 1] Baseline IL22 (A) and STAT5A (B) mRNA levels were higher in colonic tissues from VDZ non-responder (VDZ-NR) UC patients compared with responder (VDZ-R) UC patients. mRNA expression was derived from microarray data generated in moderate / severe UC patients (Gene Expression Omnibus: GSE73661). Horizontal rectangle: healthy controls (mean ± 1 SD). [Figure 2] Serum IL-22 levels at baseline were similar in VDZ-remitters and non-remitters, but remained proportionally higher in non-remitters. Serum IL-22 was assessed in patients with moderate-to-severe CD. (A) IL-22 levels at baseline and week 10 in VDZ-remitters and non-remitters. IL-22 levels were censored at the lower limit of quantification (2.7 pg / ml). (B) Fold change in IL-22 from baseline to week 10. [Figure 3] 1 provides a schematic diagram of the study described in Example 1. [Figure 4A] The results of the mouse colitis study described in Example 1 are presented graphically, in which colon weight, diarrhea score, and histopathology score were quantified for healthy mice (normal), mice injected with a negative IgG control, anti-MAdCAM antibody, anti-p40 antibody, and both anti-MAdCAM antibody and anti-p40 antibody. [Figure 4B] The results of the mouse colitis study described in Example 1 are presented graphically, in which colon weight, diarrhea score, and histopathology score were quantified for healthy mice (normal), mice injected with a negative IgG control, anti-MAdCAM antibody, anti-p40 antibody, and both anti-MAdCAM antibody and anti-p40 antibody. [Figure 4C]The results of the mouse colitis study described in Example 1 are presented graphically, in which colon weight, diarrhea score, and histopathology score were quantified for healthy mice (normal), mice injected with a negative IgG control, anti-MAdCAM antibody, anti-p40 antibody, and both anti-MAdCAM antibody and anti-p40 antibody. [Figure 5] (A) Graphical representation of the results of the study in Example 1 showing CD3% in the lamina propria (LP) and epithelium (EL) for the IgG-injected group (negative control), the anti-MAdCAM antibody-injected group, the anti-p40 antibody-injected group, and the anti-MAdCAM antibody and anti-p40 antibody-injected group. (B) Correlation between the percentage of CD3 and total inflammatory cell infiltration. [Figure 6] Panels A and B graphically depict the results showing the levels of neutrophils (Figure 6A) and macrophages (Figure 6B) in the lamina propria 28 days after administration of IgG, anti-MAdCAM-1 antibody, anti-p40 antibody, or a combination of anti-MAdCAM-1 and anti-p40 antibodies. [Figure 7] 1 is a graphical representation of a diagram showing the synergistic effect of gene expression using a combination of anti-MAdCAM-1 and anti-p40 therapy. [Figure 8] Figures A-C show graphs of the results of administering a vehicle control (negative control), anti-MAdCAM-1 antibody (MAdCAM-1), anti-p40 antibody (p40), or a combination of anti-MAdCAM-1 and anti-p40 antibodies (combo) in a mouse model of colitis. Normal mice were used for comparison. Figure A shows the body weight (g) of mice on day 21, Figure B shows the body weight of mice on day 28, and Figure C shows the diarrhea score of mice on day 21. [Figure 9A] Figures 9A, 9B, and 9C show the results of administering a vehicle control (negative control), anti-MAdCAM-1 antibody (MAdCAM-1), anti-p40 antibody (p40), or a combination of anti-MAdCAM-1 and anti-p40 antibodies (combo) in a mouse model of colitis. Figures 9A, 9B, and 9C show the levels of CD3+ T cells, MPO+ neutrophils, and CD68+ macrophages, respectively, in the colonic mucosa of each tested group and a normal control. [Figure 9B]Figures 9A, 9B, and 9C show the results of administering a vehicle control (negative control), anti-MAdCAM-1 antibody (MAdCAM-1), anti-p40 antibody (p40), or a combination of anti-MAdCAM-1 and anti-p40 antibodies (combo) in a mouse model of colitis. Figures 9A, 9B, and 9C show the levels of CD3+ T cells, MPO+ neutrophils, and CD68+ macrophages, respectively, in the colonic mucosa of each tested group and a normal control. [Figure 9C] Figures 9A, 9B, and 9C show the results of administering a vehicle control (negative control), anti-MAdCAM-1 antibody (MAdCAM-1), anti-p40 antibody (p40), or a combination of anti-MAdCAM-1 and anti-p40 antibodies (combo) in a mouse model of colitis. Figures 9A, 9B, and 9C show the levels of CD3+ T cells, MPO+ neutrophils, and CD68+ macrophages, respectively, in the colonic mucosa of each tested group and a normal control. DETAILED DESCRIPTION OF THE INVENTION

[0048] definition In order that the present invention may be more readily understood, certain terms are first defined. In addition, Whenever a value or range of values ​​for a parameter is listed, values ​​intermediate the listed values ​​and It should be noted that ranges are also intended to be part of the present invention.

[0049] The cell surface molecule "α4β7 integrin" or "α4β7" (interchangeably throughout) used) is the α4 chain (CD49D, ITGA4, OMIM 192975, human Gene I D3676) and β7 chain (ITGB7, OMIM 147559, human GeneID36 95) is a heterodimer of the human α4-integrin and β7-integrin genes. (GenBank (National Center for Biotechnology Information) Neurology Information, Bethesda, MD) RefSeq Accession Number Session numbers NM_000885 and NM_000889) are used to identify B and T lymphocytes. It is expressed by lymphocytes, especially memory CD4+ lymphocytes. α4β7 can exist in either a resting or active state. Ligands for α4β7 include: Vascular cell adhesion molecule (VCAM), fibronectin, and mucosal addressin (MAdCAM) (e.g., MAdCAM-1).

[0050] As used herein, "inhibitors of α4β7 integrin" or "α4β7 inhibitors" refers to The drug "can bind to a ligand, e.g., MAdCAM, VCAM, or α4 to fibronectin." Inhibits the binding of β7 integrin. Inhibits the binding of α4β7 integrin to MAdCAM. α4β7 inhibitors that inhibit α4 integrin, β7 integrin, or α4 integrin and β7 integrin. To inhibit α4β7 integrin activity, MAdCAM-α4β7 integrin binding was Administering a polypeptide that inhibits α4β7 integrin is an "anti-α4β7 integrin therapy." In certain embodiments, polypeptides that inhibit MAdCAM-α4β7 integrin binding anti-α4β7 integrin antibodies, e.g., antibodies that bind to either α4 or β7 only in the presence of the other. an antibody that binds to the antibody, such as vedolizumab, or a related antibody, or an antigen-binding fragment thereof. In some embodiments, the α4β7 inhibitor binds to α4β7 integrin and inhibits MA Blocking the interaction of α4β7 integrin with dCAM-1 allows memory T lymphocytes to bind to the endothelium. inhibits migration of inflammatory gastrointestinal agents through the GI tract into the inflamed gastrointestinal parenchyma.

[0051] As used herein, an antibody or antibody having "binding specificity for the α4β7 complex" The antigen-binding fragment binds to α4β7 but not to α4β1 or αEB7. Vedolizumab is an example of an antibody with binding specificity for the α4β7 complex. .

[0052] As used herein, "anti-α4β7 antibody" or "anti-α4β7 integrin antibody" refers to an antibody that " refers to an antibody that specifically binds to the α4β7 integrin. In one embodiment, anti-α4β 7 antibodies block or inhibit binding of α4β7 integrin to one or more of its ligands In one embodiment, the anti-α4β7 antibody binds to α4β7 but not to α4β1 or αE In one embodiment, the anti-α4β7 antibody is vedolizumab.

[0053] The term "antibody" broadly refers to a group of four polypeptides interconnected by disulfide bonds. It refers to an immunoglobulin molecule consisting of a single antibody chain, two heavy (H) chains, and two light (L) chains. Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region ( The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions further The complementarity determining region is sandwiched between more conserved regions called framework regions (FR). Each VH and VH can be subdivided into regions of hypervariability called constant domains (CDRs). L is in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 , consisting of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus become.

[0054] As used herein, an "antibody fragment" or "antigen-binding fragment" of an antibody The term "antibody" refers to Fab, Fab', F(ab)2, and Fv fragments, single chain antibodies Nanobodies, functional heavy chain antibodies (nanobodies), and those that compete with intact antibodies for specific binding Any portion of an antibody (e.g., a polypeptide) having specificity for at least one desired epitope. For example, a complementary sequence having sufficient framework sequence to specifically bind to the epitope. Antigen-binding fragments are isolated portions of the antibody-binding region of a human antigen. It can be produced by enzymatic or chemical cleavage of the body.

[0055] As used herein, the term "humanized antibody" refers to an antibody that retains the antigen-binding properties of the parent antibody. These antibodies retain or substantially retain the immunogenicity of non-human immunoglobulins, but are less immunogenic in humans. refers to an antibody derived from a non-human antibody (e.g., mouse) that contains minimal sequence derived from Generally, humanized antibodies are antibodies in which residues from the hypervariable region of a recipient antibody have been replaced with residues from a non-human species (donor antibody). mouse, rat, rabbit, or other suitable host with the desired specificity, affinity, and capacity. Residues from the hypervariable regions of human immunoglobulins (e.g., human primates) are replaced by residues from the hypervariable regions of the corresponding non-human primates. In some cases, the framework regions of human immunoglobulins are Furthermore, humanized antibodies are characterized by the replacement of certain non-human (FR) residues with corresponding non-human residues. These modifications may also include residues that are not found in the recipient antibody or the donor antibody. Furthermore, humanization is performed to improve antibody performance. Generally, humanized antibodies contain at least one , which will usually include substantially all of the two variable domains, and the hypervariable loops All or substantially all of the variable loops (complementarity determining regions) are from non-human immunoglobulins. and all or substantially all of the FRs correspond to those of a human immunoglobulin sequence. Humanized antibodies may also optionally contain an immunoglobulin constant region (Fc), typically a human For further details, see Jones et al.,Nature 321:522-525(1986);Ri echmann et al., Nature 332:323-329(1988); and Presta, Curr. Op. Struct. Biol. 2:593-596(1 See 992).

[0056] As used herein, the term "IL-23 inhibitor" refers to an agent that inhibits or suppresses IL-23 activity. refers to an agent that reduces the binding of IL-23 and / or IL-23R. Alternatively, the agent may inhibit IL-23mR. These inhibitors may act to reduce levels of IL-23 or IL-23 protein. Examples of therapeutic agents include antibodies or antigen-binding fragments thereof, small molecules, and nucleic acids (e.g., mRNA, DNA, siRNA, shRNA, antisense RNA, miRNA) The IL-23 inhibitor can be, but is not limited to, an IL-23 subunit ( p19 or p40), or alternatively, both subunits. It can act on subunits, for example, overlapping or combinatorial epitopes. In some embodiments, the IL-23 inhibitor binds to p19 (e.g., an anti-p19 antibody). In another embodiment, the IL-23 inhibitor inhibits or reduces IL-23 activity. 0 (e.g., anti-p40 antibodies). The terms "p19" and "p40" are used herein. As used herein, they refer to the individual subunits that make up human IL-23. In some embodiments, the IL-23 inhibitor is an IL-23R (its subunit I). Thus, in some embodiments, The term "IL-23 inhibitor" refers to an IL-23 inhibitor that inhibits IL-23 receptors, such as p19 (e.g., anti-p19 antibodies), p40 (e.g., For example, an anti-p40 antibody), or an IL-23R (anti-IL-23R antibody). The term "antibody" encompasses agents that bind to or inhibit one or more of the following: can be.

[0057] As used herein, the term "monoclonal antibody" refers to a population of substantially homogeneous antibodies. The term "antibodies" refers to antibodies obtained from a population, i.e., the individual antibodies comprising the population are called monoclonal antibodies. identical and / or the same epitope, except for possible variants that may arise during the generation of Such variants generally exist in small amounts. In contrast to polyclonal antibody preparations, which contain different antibodies against each monoclonal antibody (group), A monoclonal antibody is one that is directed against a single determinant on an antigen. The term refers to the characterization of an antibody obtained from a substantially homogeneous population of antibodies and by any particular method. It should not be construed as requiring the production of antibodies. The monoclonal antibody is described in Kohler et al., Nature, 256:49 5 (1975), recombinant DNA technology It may be produced by Method A (for example, U.S. Patent No. 4,816,567). "Clonal antibodies" are also used herein, for example, as described in Clackson et al., Nature, 35 2:624-628 (1991) and Marks et al., J. Mol. Biol. ., 222:581-597 (1991) using the method described in It may be isolated from the larvae.

[0058] As used herein, the term "recombinant antibody" refers to an antibody expressed on a recombinant expression vector. In one embodiment, the term "antibody" refers to an antibody produced as a result of transcription and translation of a gene carried by The vector is introduced into a host cell. Alternatively, the vector can be used in a cell-free system. can.

[0059] The term "initial time" refers to the starting point used for comparison. In one embodiment, the initial time is , at a time point prior to treatment with an anti-α4β7 antibody or antigen-binding fragment thereof, e.g., at 0 Refers to the day.

[0060] Terms like "treatment" or "treating" refer to the prevention or prophylaxis of a disease or disorder. To prevent, i.e., to prevent the onset of clinical symptoms; to suppress, or inhibit, a disease or disorder and / or preventing or inhibiting the onset of clinical symptoms; and / or preventing or inhibiting the progression of a disease or disorder. a disease in a human subject, including reducing harm, i.e., causing regression of clinical symptoms. In one embodiment, treatment of IBD is achieved, and Subjects with a recognized IBD index (e.g., clinical signs of Crohn's disease or ulcerative colitis) Symptoms measured with a dosing regimen using anti-α4β7 antibody and IL-23 inhibitor See the improvement.

[0061] The term "therapeutically effective dose" refers to the treatment of a disease and its associated diseases in a patient already suffering from the disease. is defined as an amount sufficient to cure or at least partially prevent the complications of In embodiments, a therapeutically effective dose improves symptoms in a human subject with the disease and / or Eliminate or reduce complications associated with IBD (e.g., those caused by long-term steroid use) In one embodiment, the dose of the anti-α4β7 antibody and the dose of the IL-23 inhibitor are Therefore, a therapeutically effective dose is one that reduces or eliminates the Crohn's Disease Activity Index (CDAI) score. or modified CDAI from a human subject diagnosed with Crohn's disease, as defined by Crohn's disease. The dose of anti-α4β7 antibody and IL-23 inhibitor that can reduce the score to the lowest In one embodiment, a therapeutically effective dose is a dose that reduces or eliminates the Mayo score. or a score lower than that defined for ulcerative colitis in human subjects diagnosed with ulcerative colitis. The dose of the anti-α4β7 antibody and the dose of the IL-23 inhibitor can be reduced to

[0062] As used herein, the term "clinical" as used herein with respect to ulcerative colitis subjects The term "remission" refers to a perfect Mayo score of 2 points or less and a perfect Mayo score of more than 1 point. does not refer to individual subscores. Crohn's disease "clinical remission" is defined as a C score of 150 points or less. Refers to the DAI score.

[0063] The term "clinical response" as used herein with respect to ulcerative colitis subjects means a response of 1 point A reduction in the rectal bleeding subscore of ≥1 point or an absolute rectal bleeding score of ≤1 point A complete Mayo score reduction of 3 or more points and a 30% (or complete) reduction from baseline If the full Mayo score was not performed at the visit, a score of 2 points or more from baseline and 25 points or more from baseline % or greater). A "clinical response" was defined as a reduction of 70 or more points in the CDAI score from the start of treatment (week 0). Point.

[0064] As used herein, a "non-remitter," "remitter," or "vedolizumab non-remitter" refers to a patient who: The term "subject" refers to a subject with an autoimmune disease and / or IBD (e.g., Crohn's disease or ulcerative colitis). are used interchangeably to refer to the subset of patients with α4β7 inhibitors (inflammatory bowel disease, inflammatory bowel disease, and inflammatory bowel disease), first and second induction doses of anti-α4β7 antibodies (e.g., vedolizumab), or or third induction dose and early in therapy, e.g., vedolizumab therapy (e.g., , about 3 or 4 weeks after the second or third challenge dose) show signs of non-remission. For example, Patients were randomly assigned to receive an anti-α4β7 antibody (e.g., vedolizumab) at 0 and 2 weeks, or The patient may receive the first dose at 3-6 weeks, for example, about 3-4 weeks after the second dose. For example, 5 or 6 weeks after starting treatment with an anti-α4β7 antibody (e.g., vedolizumab) or about 3 to 8 weeks after the third dose, e.g., 10 or 14 weeks after initiation of treatment. A patient may show clinical response to steroids but not clinical remission. Signs of non-remission include: failure to achieve clinical remission measures and / or measures described herein It may also include.

[0065] As used herein, the term "about" is used synonymously with the term "approximately." For explanatory purposes, use of the term "about" refers to a range slightly outside the quoted value, i.e., ± The value indicates 5%.

[0066] Therapeutic Uses and Methods Use a combination therapy that includes an α4β7 inhibitor, e.g., an anti-α4β7 antibody, and an IL-23 inhibitor. and methods for treating a human patient in need thereof are provided herein. The method is based, at least in part, on the discoveries described in the Examples below. Anti-α4β7 antibodies, such as α4β7 antibodies, or their antigen-binding portions, promote the recruitment of α4β7 T cells to the lamina propria of the colon. Blocking transport reduces inflammation associated with IBD, e.g., Crohn's disease. IL-23 inhibitors, such as anti-IL23 antibodies, or antigen-binding portions thereof, inhibit the natural immunity of the lamina propria. Reduced inflammation in human subjects with Crohn's disease through blockade of immune cell-derived IL-23 To make.

[0067] Suppression or downregulation of the gut adaptive and innate immune systems by vedolizumab and IL-23 inhibitors The two drugs complement each other to treat intestinal inflammation in subjects with IBD. Provides improvements above.

[0068] "Combination therapy" in the context of administration refers to the use of two or more therapies, e.g., two or more drugs, e.g. For example, an α4β7 inhibitor, e.g., an anti-α4β7 antibody, or an antigen-binding portion thereof, and an IL- 23 inhibitors, such as anti-p40, anti-p19 antibodies, or anti-IL-23R antibodies, or "In combination" is also intended to mean the use of these antigen-binding fragments. The use of the term "combination therapy" does not restrict the order in which therapies are administered to a subject with a disease. The initial treatment is a two-dose regimen for patients with diseases such as inflammatory bowel disease (IBD). Before administration of eye treatment (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks , 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks) simultaneously with or after administration After (e.g., 1 minute, 45 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 o'clock) 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks Optional additional therapy may be administered for 1 week, 6 weeks, 8 weeks, or 12 weeks. It may be administered in any order with other additional treatments.

[0069] In one embodiment, the present invention provides a method of treating a human patient in need thereof, the method comprising: The method comprises administering to a human patient an α4β7 inhibitor, e.g., an anti-α4β7 antibody, e.g., a humanized anti-α4β 7Antibodies (e.g., vedolizumab), or antigen-binding portions thereof, and IL-23 inhibitors (e.g., For example, an anti-p40, anti-p19 antibody, anti-IL-23R antibody, or an antigen-binding portion thereof) In certain embodiments, the human patient is a patient suffering from, but not limited to, psoriasis or arthritis. autoimmune diseases, including arthritis (e.g., rheumatoid arthritis, psoriatic arthritis, axial spondyloarthritis, juvenile arthritis) Autoimmune diseases and / or inflammation, such as, but not limited to, ulcerative colitis or Crohn's disease In certain embodiments, the human patient has moderate to severe active inflammatory bowel disease (IBD). In another embodiment, the human patient has moderate to severe active ulcerative colitis. He has leukemia.

[0070] In one embodiment, an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab) or an antigen-binding portion thereof may be used in combination with an IL-23 inhibitor (e.g., anti-p40, anti-IL-23 inhibitor) for treatment. p19 antibody, anti-IL-23R antibody, or antigen-binding portion thereof) before other In embodiments, the anti-α4β7 antibody (e.g., vedolizumab), or antigen-binding portion thereof, is IL-23 inhibitors (e.g., anti-p40, anti-p19, or anti-IL-23R antibodies, or In yet another embodiment, the anti-α4β7 antibody (e.g., its antigen-binding site) is administered after the anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, may be administered to an IL-23 inhibitor (e.g., anti-p4 0, anti-p19 antibody, or anti-IL-23R antibody, or antigen-binding portion thereof).

[0071] The methods disclosed herein include administering an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., Bedri izumab) or an antigen-binding portion thereof, and an IL-23 inhibitor (e.g., anti-p40, anti-p19 and administering both an anti-IL-23R antibody, an anti-IL-23R antibody, or an antigen-binding portion thereof. In some embodiments, an α4β7 inhibitor, such as an anti-α4β7 antibody ( (e.g., vedolizumab), or antigen-binding portions thereof, in accordance with approved protocols. may be administered, and IL23 inhibitors (e.g., anti-p40, anti-p19 antibodies, anti-IL-23R antibodies, or antigen-binding portions thereof) when administered according to approved protocols. good.

[0072] The combination therapies disclosed herein include an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab), or an antigen-binding portion thereof, and an IL-23 inhibitor (e.g., anti-p40 Antibodies (anti-p19 antibodies, or antigen-binding portions thereof) can be delivered via various routes known in the art, including: For example, it may be administered orally, transdermally, subcutaneously, intranasally, intravenously, intramuscularly, intraocularly, or parenterally. The most suitable route of administration in any given case will depend on the particular α4β7 inhibitors, such as anti-α4β7 antibodies and IL-23 inhibitors, and e.g. Patient, formulation, method of administration (e.g., administration time), patient age, weight, body surface area, sex, treatment The dosage will depend on the severity of the disease being treated, the patient's diet, and the patient's excretion rate.

[0073] In the combination therapies disclosed herein, an IL-23 inhibitor (e.g., an anti-IL23 antibody or anti-IL23R antibody) before administration of anti-α4β7 antibody (e.g., vedolizumab). Patients in need (e.g., patients with autoimmune diseases such as arthritis or psoriasis, and / or or inflammatory diseases such as IBD. In the combination therapies disclosed herein, the IL-23 inhibitor is used in combination with an α4β7 inhibitor, e.g., an anti-α 1 minute to 1 week (e.g., 1 minute, 5 minutes, 10 minutes) after administration of a 4β7 antibody (e.g., vedolizumab) minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 o'clock Between, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 1 1 hour, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 1 9 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days The patient may be administered the drug 1 day, 6 days, or 7 days or more before the administration of the drug.

[0074] Alternatively, in the combination therapies disclosed herein, an IL-23 inhibitor (e.g., an anti-IL- 23 antibody or anti-IL23 antibody) after administration of an α4β7 inhibitor, e.g., an anti-α4β7 antibody and patients in need thereof (e.g., those with autoimmune diseases such as arthritis or psoriasis and / or can be administered to patients with inflammatory diseases such as IBD. In the combination therapy disclosed herein, the IL-23 inhibitor is used in combination with an α4β7 inhibitor, e.g. For example, 1 minute to 1 week (e.g., 1 minute, 5 minutes) after administration of an anti-α4β7 antibody (e.g., vedolizumab) minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes Minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours Hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours Hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days The patient may be administered the drug 2 days, 5 days, 6 days, or 7 days or more later.

[0075] The combination therapy disclosed herein may be administered for 1 week to 10 years (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, Week, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 1 0 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years) and / or for any given period of time necessary to treat a disease, e.g., IBD, including For example, in the combination therapies disclosed herein, an α4β7 inhibitor, e.g., an anti-α4 β7 antibodies (e.g., vedolizumab) and IL-23 inhibitors (e.g., anti-IL-23 antibodies) is administered to a patient in need thereof (e.g., a patient with arthritis or patients with autoimmune diseases such as psoriasis and / or inflammatory diseases such as IBD) for 1 week ~10 years (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 1 week, 2 weeks, 3 weeks, 4 weeks 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 1 year, 2 years, 3, 4, 5, 6, 7, 8, 9, or 10 years) can.

[0076] In some embodiments, the treatment of an autoimmune disease (e.g., arthritis or psoriasis—e.g., joint rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, axial spondyloarthritis) or chronic inflammatory diseases (e.g. A patient with IBD (e.g., IBD) has one or more symptoms of the patient's disease (e.g., 5% or more, e.g., For example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50 %, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97 The patient continues to receive the combination therapy disclosed herein until the patient's blood pressure is reduced (99%, 99%, or more). For example, an IBD patient may have one or more symptoms of their IBD that are worse than when they first started. (e.g., 5% or more, e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or more) reduction in According to the described methods, an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab ) and an IL-23 inhibitor (e.g., an anti-IL-23 antibody or an anti-IL-23R antibody). For example, an IBD patient may continue to receive a combination of It will be.

[0077] In one aspect, the present invention provides a method for treating a disease or disorder, e.g., in a human, each of which is useful. an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab) and an IL-1 administering an IL-23 inhibitor (e.g., an anti-IL-23 antibody or an anti-IL-23R antibody) to the subject The human subject may be an adult (e.g., a 20-year-old man or older), or a 20-year-old man or older, and the method of treating the disease or disorder in the subject is provided. The human subject may be an adult (e.g., 18 years of age or older), an adolescent, or a child (juvenile or toddler). In certain embodiments, the human subject is a child under the age of 18. do.

[0078] In certain embodiments, the subject is responsive to treatment of the methods according to the embodiments disclosed herein. Responders were those who achieved (1) endoscopic cure; (2) clinical response; (3) Inflammatory Bowel Disease Questionnaire ( (4) change from baseline (e.g., improvement) in IBDQ score; (5) mucosal healing; (6) Decrease in C-reactive protein, fecal lactoferrin, and alfa and one or more biomarkers selected from the group consisting of fecal calprotectin. (7) Normalization of Kerr's score; (8) Improvement of Psoriasis Area and Severity Index 75 (PASI) compared to baseline; and (8) the American College of Rheumatology. Based on the 20% response criteria (ACR20) or 28-joint Disease Activity Score (DAS28) The improvement is confirmed to have at least one of the following.

[0079] In certain embodiments, the present invention provides a method for treating moderately to severely active ulcerative colitis in a subject. The present invention provides a clinically proven, safe and effective method for administering to a subject a combination of antibodies. A responder to treatment with a combination therapy, and at a given time, e.g., Mayo endoscopic subscore of 0 or 1 by 8, 12, and 16 weeks after placement have been identified as having a statistically significant improvement in disease activity as determined by endoscopic cure. can be.

[0080] In another embodiment, the present invention provides a method for treating moderately to severely active ulcerative colitis in a subject. The present invention provides a clinically proven, safe, and clinically proven effective method for administering α4β7 inhibitors to a subject. anti-α4β7 antibodies (e.g., vedolizumab) and IL-23 inhibitors (e.g., , anti-IL-23 antibody) and are candidates for treatment with combination therapy. By a given time point, e.g., week 8, week 12, week 16, there is a statistically significant difference in disease activity. Significant improvement (as determined by an Ulcerative Colitis Endoscopic Index of Severity (UCEIS) score of 4 or less) It has been confirmed that it has.

[0081] In some embodiments, the present invention provides a method for treating an inflammatory disorder (e.g., moderate to severe activity) in a subject. The present invention provides a method for treating IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21, IL Treatment with an -23 inhibitor also results in increased activity of an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., In certain embodiments, the non-responders are those who are non-responders to treatment with cerebrospinal fluid (e.g., cerebrospinal fluid, vedolizumab). Responders or non-responders may be treated with an α4β7 inhibitor, e.g., an anti-α4β7 inhibitor, as described herein. refractory to β7 antibody treatment and / or have elevated IL22 or STAT5 Other indicia of non-response may be measured using clinical response measures and / or the measures described herein. For example, the patient may receive a small dose of an α4β7 inhibitor, for example, at 0 and 2 weeks. At least two induction doses, and then characterized as non-responders after approximately 3-4 weeks After a period of time, patients do not show significant improvement in symptoms and / or achieve clinical response without clinical remission. Non-responder patients may be treated with α4β7 inhibitors (e.g., IL-23 inhibitors) in combination with IL-23 inhibitors. For example, vedolizumab 300 mg every 4 or 8 weeks, or every 2, 3, or 4 weeks Combination therapy may be continued for, for example, 4 to 20 weeks, 4 to 14 weeks, or The treatment may last for 6 to 12 weeks. In some embodiments, the IL-23 inhibitor is administered as described herein. As shown, Itgal (αL chain / CD11a / LFA-1A), Itgb2 (β2 Integrin chain / CD18), Itgax (αX chain / CD11c), Itga3, Itg a9, Itgb1bk, Il21r, Il12rb1, Il12a, IL2ra, IL1 0ra, Il17re, Il34, Il18rap, Il1rl1, Il1b, Il1r 2, Il3ra, Il1f9, Il23a, Iltifb, Il6, Il18bp, Il 1a, Il15, Il1r1, Stat4, Stat2, and Cd3g. At least one gene selected is altered (e.g., upregulated or downregulated). and / or until at least one of the genes is modified. At least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least At least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks maintained for at least 11 weeks, at least 12 weeks, or longer. In embodiments, the IL-23 inhibitor inhibits integrin-beta-2 (Itgb2 (β2 integrin) In some embodiments, the antibody is administered until downregulation of phosphochain / CD18 is achieved. IL-23 inhibitors are effective until downregulation of IL1B (interleukin 1β) is achieved. In some embodiments, the IL-23 inhibitor is administered to achieve downregulation of IL23a. Downregulation can be achieved by administering an α4β7 inhibitor, such as an anti-α4β7 antibody (e.g., For example, 5%, 1% and 2% of patients treated with IL-23 inhibitors (e.g., vedolizumab) were at risk of developing IL-23-associated 0%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 6 0%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or Downregulation can be achieved by α4β7 inhibitors, such as anti-α4β7 antibodies (e.g., Compared with before combination therapy with an IL-23 inhibitor (dolizumab), the 5%, 5%~20%, 5%~15%, 5%~10%, 10%~20%, 10%~15%, 10%~30%, 15%~40%, 20%~50%, 25%~60%, 30%~70%, Or, it may be 40% to 100% or more. In some embodiments, patients receiving combination therapy The authors suggest that once a patient achieves remission, they may consider administering an α4β7 inhibitor (e.g., an anti-α4β7 antibody, e.g., vedolizumab) monotherapy.

[0082] In certain embodiments, the combination therapy described herein is a combination therapy, e.g., By the 8th week, a reduction of 30% or more and 3 or more points in the Mayo score from the start of treatment, and a rectal bleeding subscore of 1 point or more from the start or 0 or 1. For Crohn's disease or ulcerative colitis, as determined by a reduction in bleeding subscore A human subject with IBD achieving a clinical response as defined herein is provided.

[0083] In certain embodiments, the present invention provides a method for treating IBD, e.g., moderate to severe IBD, in a human subject. Clinically proven to treat active ulcerative colitis or moderately to severely active Crohn's disease provides a safe, clinically proven, and effective method for the treatment of The identified subject is administered the combination therapy described herein.

[0084] In certain embodiments, the combination therapies described herein are used to treat Crohn's disease or ulcerative colitis. providing a human subject with IBD that achieves clinical remission as defined herein; Subjects identified as non-remitters to initial treatment are administered the combination therapy described herein. .

[0085] In certain embodiments, the subject receiving the combination therapy of the present invention is also receiving an immunomodulatory agent, such as TNF-α For treatment, e.g., initial treatment, with Inadequate response, loss of response, or intolerance to In some embodiments, the subject to be administered the combination therapy of the present invention , initial treatment with an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab) There may be subsequent insufficient response or no remission. The subject receiving the combination therapy of the present invention is one who has not responded adequately to corticosteroid therapy. have not responded to, or have been dependent on, Patients must be taking at least one corticosteroid (e.g., prednisolone) for inflammatory bowel disease. may have received prior treatment with steroids (e.g., cyclosporine). The response was equivalent to oral prednisone 30 mg given for 2 weeks or intravenously once daily for 1 week. persistence despite a history of at least one 4-week induction regimen that included Refers to the signs and symptoms of ongoing active disease. Loss of response to corticosteroids is considered Attempt to taper luteosteroids to a dose below the equivalent of 10 mg oral prednisone daily Corticosteroid intolerance is associated with Cushing's syndrome, osteopenia / osteoporosis, History of steroid use over time, including history of idiopathic steroid use, hyperglycemia, insomnia, and / or infections. Potential complications may include venous thromboembolism, fragility fractures, or infection.

[0086] Immunomodulators include, for example, oral azathioprine, 6-mercaptopurine, or methotrexate. An inadequate response to an immunomodulatory agent is defined as at least one 8-week period of regimen or oral azathioprine, 6-mercaptopurine, or methotrexate Refers to persistent signs and symptoms of active disease despite a known history. Tolerance includes, but is not limited to, nausea / vomiting, abdominal pain, pancreatitis, abnormal LFTs, and lymphopenia TPMT gene mutation and / or infection.

[0087] In one embodiment, the subject has an inadequate or inadequate response to treatment with a TNFα antagonist. have lost response to or been intolerant to TN F-α antagonists are, for example, drugs that suppress the biological activity of TNF-α, and are preferably Preferably, TNF-α, e.g., a monoclonal antibody, e.g., REMICADE (Invitrogen) fliximab), HUMIRA (adalimumab), CIMZIA (certolizumab pegol) ), SIMPONI (golimumab), or Fc fusion proteins, e.g., ENBREL (etanercept). Inadequate response to TNF-α antagonists is At least one 4-week induction regimen of infliximab 5 mg / kg IV was administered. 2 doses at 2-week intervals; adalimumab 80 mg subcutaneously once, followed by at least or certolizumab pegylated 40 mg once every 2 weeks; or certolizumab pegylated 40 mg once every 2 weeks 400 mg subcutaneously twice daily for signs and symptoms of persistent active disease despite a history of Loss of response to TNF-α antagonists refers to the lack of maintenance after previous clinical benefit. This refers to the recurrence of symptoms during treatment. Intolerance to TNF-α antagonists can be caused by infusion-related reactions, demyelination, , congestive heart failure, and / or infection.

[0088] In one embodiment, the disease that can be suitably treated with the combination therapy described herein is rectal Inflammatory bowel disease (IBD), e.g., ulcerative colitis, obtained after colectomy and ileoanal anastomosis associated with Crohn's disease, ileitis, celiac disease, non-tropical sprue, and seronegative arthropathy. enteropathy, microscopic or collagenous colitis, eosinophilic gastroenteritis, or pouchitis. In embodiments, the inflammatory bowel disease is Crohn's disease or ulcerative colitis. Ulcerative colitis is The treatment may be for moderate to severe active ulcerative colitis. It may result in mucosal healing in patients with colitis. Treatment may also improve corticosteroid activity by the patient. This may result in the reduction, elimination, or reduction and elimination of steroid use.

[0089] In other embodiments, the present invention relates to the treatment of autoimmune diseases (e.g., arthritis or psoriasis, e.g., rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, or axial spondyloarthritis), or inflammation Intestinal disease (e.g., ulcerative colitis or Crohn's disease), e.g., moderate to severe active ulcerative colitis ulcerative colitis; or moderately to severely active Crohn's disease), The use of a combination of an IL-23 inhibitor and an IL-23 inhibitor is also provided, as described herein. Thus, use is limited to patients characterized as non-responders or non-remitters to α4β7 inhibitors. In some embodiments, the method comprises administering to the subject an α4β7 inhibitor and an IL-23 inhibitor. For example, the α4β7 inhibitor is an anti-α4β7 antibody, such as vedolizumab. In some embodiments, vedolizumab is administered according to the dosing regimens described herein. .

[0090] In some embodiments, the patient is treated with an α4β7 inhibitor (e.g., an anti-α4β7 antibody, e.g., Non-responders or In some embodiments, the patient is treated with an α4β7 inhibitor (e.g., For example, at the start of or 6 weeks after the start of treatment with an anti-α4β7 antibody, e.g., vedolizumab Eyes have been characterized as having elevated levels of IL-22 in serum. In this study, patients were randomly assigned to receive vedolizumab at the start of treatment or 6 weeks after starting treatment. It has been characterized as having elevated levels of IL-22, and serum IL-22 levels are There was no decrease or 2% decrease between the start and 10th week, between the start and 6th week, or between 6th and 10th week. In some embodiments, the patient's Subjects had elevated levels of IL-22 and / or STAT5A compared to control levels. It can be characterized as:

[0091] In some embodiments, the characterization is performed prior to treatment with an α4β7 inhibitor. In some embodiments, the characterization is performed after initial treatment with an α4β7 inhibitor. In one embodiment, the IL-23 inhibitor is administered simultaneously with the α4β7 inhibitor. In some cases, an IL-23 inhibitor is administered after initial treatment with an α4β7 inhibitor. In this embodiment, the IL-23 inhibitor is administered for 2 weeks to 6 months, e.g., 2 to 4 weeks, 2 to 6 weeks 2-8 weeks, 4-6 weeks, 4-8 weeks, 4-10 weeks, 1-2 months, 1-3 months, 2 ~3 months, 2-4 months, 3-4 months, 3-5 months, 4-5 months, 4-6 months, or 5- In some embodiments, the IL-23 inhibitor is administered for 6 months until clinical remission is achieved. In certain embodiments, the IL-23 inhibitor is administered until clinical remission is achieved. 1 to 3 months after (e.g., 4 to 6 weeks, 4 to 8 weeks, 4 to 10 weeks, 6 to 8 weeks, 6 to 1 In some embodiments, the therapeutic agent is administered for up to 0 weeks, 6-12 weeks, or 2-3 months. IL-23 inhibitors target specific integrin chain genes or specific cytokine genes. In some embodiments, up to at least one selected gene is administered. The three inhibitors are Itgal (αL chain / CD11a / LFA-1A), Itgb2 (β2 inhibitor), Tegrin chain / CD18), Itgax (αX chain / CD11c), Itga3, Itga9 , Itgb1bk, Il21r, Il12rb1, Il12a, IL2ra, IL10r a, Il17re, Il34, Il18rap, Il1rl1, Il1b, Il1r2, Il3ra, Il1f9, Il23a, Iltifb, Il6, Il18bp, Il1a , Il15, Il1r1, Stat4, Stat2, and Cd3g. At least one gene to be tested is altered (e.g., upregulated or In certain embodiments, an α4β7 inhibitor (e.g., Administration of an anti-α4β7 antibody (e.g., vedolizumab) may be effective in treating patients who are no longer receiving an IL-23 inhibitor. It will continue even after the service is no longer available.

[0092] As anti-α4β7 antibodies or anti-IL-23 antibodies suitable for the methods and uses described herein, Useful antibodies are identified using techniques known in the art, such as hybridoma production. Hybridomas can be prepared, for example, using a mouse system. Protocols for immunization and subsequent isolation of splenocytes for fusion are well known in the art. Fusion partners and procedures for hybridoma production are also known. When generating antibodies, the best target protein (antigen) (whole protein or its fragment) The immunization of animals can be carried out by any method known in the art. For example, Harlow and Lane, Antibodies dies:A Laboratory Manual,New York:Cold S Spring Harbor Press, 1990. Mice, rats, sheep Methods for immunizing animals such as mice, goats, pigs, cattle, and horses are well known in the art. See, e.g., Harlow and Lane, supra, and U.S. Pat. No. 5,994,444. See, e.g., 619. The desired antigen is combined with an adjuvant to stimulate the immune response. Adjuvants known in the art include complete and incomplete flow cytometry. Int-adjuvant, RIBI (muramyl dipeptide), or ISCOM (immunostimulating complex) After immunizing an animal with a desired antigen, antibody-producing immortalized cell lines are generated by immunizing the immunized After immunization, the animal is killed and the resulting antibody is prepared from cells isolated from the animal. known methods (e.g., oncogene transduction, oncogenic viral transduction, oncogenic or mutagenic Exposure to compounds, fusion with immortalized cells, e.g., myeloma cells, and inactivation of tumor suppressor genes Lymph node and / or splenic B cells are immortalized by immunoprecipitation (i.e., immunoprecipitation). See, e.g., Harlow, supra. See, e.g., and Lane. Hybridomas are selected, cloned, and further Screen for desirable characteristics, including robust growth, high antibody production, and desirable antibody properties. Human anti-PCDH17 antibodies can be screened using HuMAb-Mouse (registered trademark). The mouse may also be generated in a mouse, such as a mouse model (e.g., mouse model 1000) or a mouse model (e.g., mouse model 1000) such as a mouse model (e.g., mouse model 1000).

[0093] Antibodies or antibody fragments for molecules capable of binding to target proteins (antigens) Methods for high throughput screening of libraries are disclosed herein. Such methods can be used to identify useful affinity matured antibodies. In particular, in vitro display techniques known in the art, such as phage display, Spray, bacterial display, yeast display, mammalian cell display, ribosomal These include RNA display, mRNA display, and cDNA display. The use of phage display to isolate ligands that bind to molecules related to, e.g., Felici et al.,Biotechnol.Annual Rev.1:14 9-183,1995;Katz,Annual Rev.Biophys.Biomo l.Struct.26:27-45,1997; and Hoogenboom et al. l., Immunotechnology 4:1-20, 1998(in vitro With respect to display techniques, the disclosures of each of these are incorporated herein by reference. Randomized combinatorial peptide libraries are reviewed in Ka y,Perspect.Drug Discovery Des.2:251-268, 1995 and Kay et al.,Mol.Divers.1:139-140 , 1996 (the disclosures of each of which, insofar as they relate to the discovery of antigen-binding molecules, are incorporated herein by reference). Select for a polypeptide that binds to a cell surface antigen described in Proteins, such as multimeric proteins, are designed to function efficiently as functional molecules. It is often displayed on phage (e.g., EP0349578; EP452783 9; and EP0589877, and Chiswell and McCaffert y,Trends Biotechnol.10:80-84 1992(antigen-binding molecules Regarding the use of in vitro display techniques for the discovery of , the disclosure of which is incorporated herein by reference). In addition, functional antibody fragments Fragments, such as Fab and scFv fragments, are suitable for in vitro display. It is expressed in the mat (e.g., McCafferty et al., Nature 348:552-554,1990;Barbas et al.,Proc.Nat Acad. Sci. USA 88:7978-7982, 1991; and Clack son et al., Nature 352:624-628, 1991 (antigen binding Regarding in vitro display platforms for gene discovery, (See, e.g., pp. 111-114, the disclosures of which are incorporated herein by reference). In particular, it can be used to identify and improve the affinity of antibodies that bind to target antigens. .

[0094] In addition to in vitro display methods, computational modeling methods have been used to identify target antigen binding sites. can be used to design and identify antibodies or antibody fragments in silico For example, using computational modeling techniques, one skilled in the art can identify extracellular epitopes of a target antigen. Libraries of antibodies, molecules capable of binding specific epitopes, such as peptides, Alternatively, antibody fragments can be screened in silico.

[0095] α4β7 inhibitors, such as anti-α4β7 antibodies, or or fragments thereof, and examples of IL-23 inhibitors are provided below.

[0096] α4β7 antagonists The methods disclosed herein include administering to a subject a therapeutically effective amount of a compound selected from the group consisting of steroids, anti-inflammatory drugs ... (e.g., rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, axial spondyloarthritis), and / or or have an inflammatory disease, such as IBD (e.g., Crohn's disease or ulcerative colitis). The subject is administered an α4β7 inhibitor, e.g., an anti-α4β7 antibody, or an antigen-binding portion thereof, and an IL-1 inhibitor. This includes administering both an IL-23 inhibitor, such as an anti-IL-23 antibody, and a IL-23 inhibitor, such as an anti-IL-23 antibody.

[0097] The various polypeptides may be used in combination with the anti-α4β7 antibodies, anti-MAdCAM antibodies, soluble integrin subunits (including fusion proteins such as Fc fusions), and soluble M α4β7 integrin, including AdCAM (including fusion proteins such as Fc fusions) MAdCAM in primates can be inhibited from binding to MAdCAM, for example, as disclosed in the PCT publication. No. WO 96 / 24673, the entire teachings of which are incorporated herein by reference. Inhibits MAdCAM-α4β7 integrin binding and is used in accordance with the present invention. Polypeptides capable of synthesizing nucleotides of the present invention are described, for example, in U.S. Pat. No. 7,803,904. As such, anti-MAdCAM antibodies (e.g., U.S. Pat. No. 8,277,808, PF-00 547659, SHP647, ontamalimab, or any of the compounds described in WO2005 / 067620 fusion proteins, such as Fc fusions, containing soluble integrin subunits; complexes containing soluble integrin subunits (e.g., α4 and / or β7, This may be a protein lacking a transmembrane domain or lacking both a transmembrane and an intracellular domain), e.g. Soluble α4 integrin, soluble β7 integrin, or soluble α4β7 integrin complexes; and fusion proteins containing MAdCAM, such as MAdCAM-Fc chimeras. Soluble MAdCAM (e.g., lacking the transmembrane domain or containing the transmembrane domain) and lacking the intracellular domain).

[0098] The present invention relies, at least in part, on anti-α4β7 antibodies or antigen-binding portions thereof, 1) binds to α4β7 integrin in vitro and / or in vivo (2) The ability or function of α4β7 integrin, for example, (a) binding function (e.g., For example, α4β7 integrin interacts with MAdCAM (e.g., MAdCAM-1), fibronectin, and (b) the ability to bind to VEGF-1 and / or VCAM-1; and / or (c) the ability to bind to VEGF-1 and / or VCAM-1 in tissues. Leukocyte infiltration, including recruitment and / or accumulation of blood cells (e.g., lymphocytes in the intestinal mucosa) In one embodiment, the antibodies used herein may modulate the ability of: The α4β7 integrin can bind to the ligand (e.g., MAdC one or more of the following: AM (e.g., MAdCAM-1), VCAM-1, fibronectin) This inhibits the binding of leukocytes to the tissues, thereby reducing leukocyte infiltration (recruitment and In another embodiment, the anti-α4 The β7 antibody can bind to the α4β7 integrin and detect when the α4β7 integrin binds to a ligand (e.g., For example, MAdCAM (e.g., MAdCAM-1), VCAM-1, and fibronectin By selectively inhibiting the binding of one or more of these proteins to the leukocytes in the tissue, Such anti-α4β7 antibodies can inhibit the inflammatory process (including the recruitment and / or accumulation of leukocytes within the esophagus). reported that cells bearing α4β7 integrin are expressed in gut-associated tissues, lymphoid organs, or leukocytes (particularly In particular, cells adhere to vascular endothelial cells in mucosal tissues containing lymphocytes such as T cells or B cells. This can be inhibited in vitro and / or in vivo. In some embodiments, the anti-α4β7 antibody used herein is an antibody that binds α4β7 and MAdCAM (e.g., M It can inhibit the interaction with AdCAM-1 and / or fibronectin. In embodiments, the anti-α4β7 antibodies used herein are directed to, for example, VCAM and α4β7 MAdCAM (e.g., MAdCAM-1) without inhibiting the interaction of The interaction with and / or selectivity for fibronectin may be inhibited.

[0099] Thus, the anti-α4β7 antibodies used in the methods disclosed herein are capable of binding to the α4β7 integrin. modulates (e.g., inhibits) the binding function and / or infiltration function of leukocytes (e.g., lymphocytes, monocytes) For example, it can be used to inhibit (reduce or prevent) α4β7 integrin. Humanized immunoglobulins that inhibit the binding of a steroid hormone to its ligand (i.e., one or more ligands). The agonist induces the proliferation of tissues (including the recruitment and / or accumulation of leukocytes in tissues), particularly the molecular MAd Leukocytes (e.g., lymphocytes, monocytes) of tissues expressing CAMs (e.g., MAdCAM-1) The compounds can be administered according to the method in the treatment of diseases associated with globular infiltration.

[0100] The anti-α4β7 antibodies used in the methods provided herein are, in certain embodiments, α4 epitopes on the α4 chain, the β7 chain, or the combination formed by the association of the α4 chain with the β7 chain In one embodiment, the antibody is specific for the α4β7 integrin complex. For example, an antibody binds to a combination epitope on the α4β7 complex, but the chains It does not bind to epitopes on the α4 or β7 chains unless they are associated with each other. In this study, anti-α4β7 antibodies bind to both the α4 and β7 integrin chains. , and therefore specific for the α4β7 integrin complex. Specific such antibodies may, for example, bind to α4β7 but not to α4β1, and / or α E In another embodiment, the anti-α4β7 antibody is incompatible with an Act-1 antibody. bind to the same or substantially the same epitope (Lazarovits, AI et al., J. Immunol., 133(4):1857-1862(1984), Sc hweighoffer et al., J. Immunol., 151(2):717 -729,1993;Bednarczyk et al.,J.Biol.Chem. ,269(11):8348-8354,1994). Accession number PTA-36 63, American Type Culture Collection (US 2 University Boulevard, Manassas, Virginia 0110-2209 10801) and Millennium Pharmaceuticals, Inc. (40 Lansdowne Street, Cambridge, Massachusetts, USA 02139) Next, we isolated mouse ACT-1 hybridoma cells that produce mouse Act-1 monoclonal antibodies. The cell line was deposited under the provisions of the Budapest Treaty on August 22, 2001. The anti-α4β7 antibody is provided in U.S. Patent Application Publication No. 2010 / 0254975. In another embodiment, the anti-α4β7 antibody is a human antibody or α4β7 binding protein that utilizes the DR. 4β7 integrin antibodies include AMG181 (Abrilumab, specific for α4β7, e.g., See U.S. Patent No. 8,444,981), etrolizumab (β7 specific, FIB504 or or humanized derivatives (e.g., Fong et al., U.S. Pat. No. 7,528,236 ), CAS1044758-60-2, KEGG D09901, PubChem 12 4490613; see, e.g., U.S. Patent No. 7,528,236), natalizumab (α4 specific Heterologous, humanized MAb 21.6, TYSABRI®, CAS 189261- 10-7, KEGG D06886, PubChem 49661786; e.g., US (See Japanese Patent No. 5840299), any of the above-mentioned related antibodies, any of the above-mentioned antigen-binding Anti-α4β7 integrin antibody was used. The treatment method is described in Publication No. US2005 / 0095238 and Publication No. WO2012151 248 and WO2012 / 151247.

[0101] An effective amount of an α4β7 inhibitor, e.g., an anti-α4β7 antibody, is used to treat such diseases. For example, the gastrointestinal tract (including gut-associated endothelium), other mucosal tissues, or tissues that express the molecule MAdCAM (e.g., MAdCAM-1) (e.g., gut-associated tissues) , e.g., the venules of the lamina propria of the small and large intestine; and the mammary gland (e.g., lactating mammary gland) Inflammatory diseases, including diseases associated with leukocyte infiltration, can be treated according to this method. Similarly, cells expressing MAdCAM (e.g., MAdCAM-1) (e.g., endothelial cells) Individuals with diseases associated with leukocyte infiltration of tissues as a result of leukocyte binding to the erythrocyte (cells) are Treatment can be performed according to the invention.

[0102] In certain embodiments, an α4β7 inhibitor, e.g., an anti-α4β7 antibody, or an antigen-binding The fragment is a humanized anti-α4β7 antibody, or an antigen-binding fragment thereof, i.e., and is an IgG1 antibody; and has a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3 a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO: 2, and a DR2 domain; The CDR3 domain set forth in SEQ ID NO: 8, the CDR2 domain set forth in SEQ ID NO: 7, and the sequence It comprises a light chain variable region comprising the CDR1 domain set forth in number 6.

[0103] In certain embodiments, the anti-α4β7 antibody, or antigen-binding fragment thereof, is selected from the group consisting of SEQ ID NO: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:1 and the amino acid sequence of SEQ ID NO:5 It is a humanized anti-α4β7 antibody containing a light chain variable domain comprising:

[0104] In a specific embodiment, the anti-α4β7 antibody is vedolizumab.

[0105] The anti-α4β7 antibody, for example, a humanized anti-α4β7 antibody, is vedolizumab, or vedolizumab. In some embodiments, the human patient receives an anti-α4 antibody at week 0. an initial dose of 300 mg of a β7 antibody, e.g., a humanized anti-α4β7 antibody, followed by an anti-α a second dose of 300 mg of a 4β7 antibody, e.g., a humanized anti-α4β7 antibody, followed by a second dose of 300 mg of a humanized anti-α4β7 antibody at week 6; A third dose of 300 mg of an anti-α4β7 antibody, eg, a humanized anti-α4β7 antibody, is administered. The method further includes administering to the human patient 300 mg of an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody. The method may include administering the β7 antibody every 8 weeks starting 8 weeks after the third dose. In embodiments, the methods described herein are performed in clinical trials in human patients with Crohn's disease or ulcerative colitis. If the patient does not show satisfactory improvement, e.g., clinical remission, 300 mg of anti-α4β7 antibody, e.g., The administration of a human anti-α4β7 antibody to a human patient may include administering the antibody every four weeks. So, the method is, for example, 300 mg of anti-α4β7 every 4 weeks starting 8 weeks after the third dose. The method includes intravenously administering an antibody, such as a humanized anti-α4β7 antibody, to a human patient. In certain embodiments, the method comprises administering to a human patient every two weeks starting eight weeks after the third dose. , 108 mg of an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody. In certain embodiments, the method includes administering an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody, at week 0. an initial dose of 300 mg, followed by an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody, at week 2. and administering a second dose of 300 mg of the antibody, followed at week 6 by administering an anti-α4β7 antibody, e.g., A third dose of 108 mg of humanized anti-α4β7 antibody, followed by 108 mg every 2 weeks thereafter In a particular embodiment, the 108 mg dose is administered subcutaneously. In certain embodiments, the 108 mg dose is self-administered.

[0106] An anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody, is vedolizumab or a vedolizumab-linked In some embodiments, the antibody has a binding region, and the juvenile (pediatric) human patient is For example, a young human patient can be treated according to the methods provided in An initial dose of 200 mg of an antibody, for example, a humanized anti-α4β7 antibody, followed by administration of the antibody two weeks after the initial dose. A second dose of 200 mg and a third dose of 200 mg of antibody administered 6 weeks after the first dose The method further includes administering a fourth dose of 200 mg 14 weeks after the first dose. The method may further comprise administering subsequent doses of 200 mg every 8 weeks thereafter. In an alternative embodiment, the young human patient may also receive an anti-α4β7 antibody, e.g. For example, the initial administration of 100 mg of humanized anti-α4β7 antibody, followed by 100 mg of antibody two weeks after the initial administration, A second dose of 100 mg of antibody will be administered, and a third dose of 100 mg of antibody will be administered six weeks after the first dose. The method further includes administering a fourth dose of 200 mg 14 weeks after the first dose. The method may further comprise administering a fifth dose of 200 mg every 8 weeks after the fourth dose and In another embodiment, the young human patient may also be administered an anti-α4β7 antibody, e.g., The first dose was 150 mg of the methylated anti-α4β7 antibody, followed by two doses of 150 mg of the antibody two weeks after the first dose. The patient will receive a first dose of 150 mg of antibody, followed by a third dose of 150 mg six weeks after the first dose. The method further comprises administering a fourth dose of 150 mg 14 weeks after the first dose. The method may further include a fourth dose of 300 mg 14 weeks after the first dose. This includes a fifth and subsequent doses of 150 mg or 300 mg every 8 weeks after the fourth dose. In certain embodiments, the young human patient may be administered an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody. First dose of α4β7 antibody 300 mg, followed by a second dose of 300 mg of antibody 2 weeks after the first dose and a third dose of 300 mg of antibody six weeks after the first dose. and a fourth dose of 300 mg 14 weeks after the first dose. A fifth and subsequent dose of 300 mg every 8 weeks after the first dose may also be included. In this embodiment, a young human patient receives 200 mg of an anti-α4β7 antibody, e.g., a humanized anti-α4β7 antibody. The first dose, the second dose of 200 mg of antibody, was administered two weeks after the first dose, and the second dose of 200 mg of antibody was administered 6 weeks after the first dose. A third dose of 108 mg of antibody was administered 1 week later, followed by 108 mg of antibody every 2, 3, or 4 weeks. Subsequent doses of g are administered subcutaneously. In some embodiments, children weighing less than 30 kg are given Patients are administered higher doses than pediatric patients weighing over 30 kg. See, e.g., PCT Publication No. WO2018 / 200818 (incorporated by reference in its entirety). is described in.

[0107] In particular, the methods disclosed herein involve the use of the anti-α4β7 antibody vedolizumab, or Vedolizumab is also available under the trade name ENTYVIO. (Registered Trademark) (Pharmaceuticals, Inc.). Vedolizumab have developed a mutated human IgG1 framework region and a murine antibody Act-1 (U.S. Patent No. 7,447,447). and antigen binding proteins from the nucleotide sequences described in US Pat. No. 6,147,851 (incorporated herein by reference). It is a humanized antibody containing mixed CDRs.

[0108] Vedolizumab specifically binds to α4β7 integrin and inhibits adhesion of α4β7 integrin. Interaction of membrane addressin cell adhesion molecule-1 (MAdCAM-1) with fibronectin Blocks the migration of memory T lymphocytes across the endothelium into inflamed gastrointestinal parenchyma Vedolizumab does not bind to or inhibits the α4β1 and αEβ7 integrins. Inhibits the function and interaction of α4 integrin with vascular cell adhesion molecule-1 (VCAM-1) Does not neutralize.

[0109] α4β7 integrin is a distinct subset of memory T lymphocytes that preferentially migrate to the gastrointestinal tract MAdCAM-1 is expressed on the surface of intestinal epithelial cells and transports lymphoid tissue to the intestine. MAd of α4β7 integrin plays an important role in T lymphocyte homing. Interaction with CAM-1 may contribute to the development of mucosal inflammation, such as the chronic inflammation that is the hallmark of ulcerative colitis and Crohn's disease. Vedolizumab is a promising treatment for Crohn's disease and ulcerative colitis. Inflammatory bowel disease including inflammatory bowel disease, HIV, pouchitis including chronic pouchitis, fistulizing Crohn's disease, graft-versus-host disease These diseases may be used to treat cereals containing glutamic acid, ... Combination therapy as described herein is contemplated.

[0110] The heavy chain variable region of vedolizumab is provided herein as SEQ ID NO: 1. The light chain variable region of vedolizumab is provided herein as SEQ ID NO: 5. A heavy chain variable region comprising CDR1 of SEQ ID NO: 2, CDR2 of SEQ ID NO: 3, and CDR3 of SEQ ID NO: 4. Vedolizumab contains CDR1 of SEQ ID NO: 6, CDR2 of SEQ ID NO: 7, and CDR3 of SEQ ID NO: The light chain variable region contains the CDR3 of SEQ ID NO: 8. The sequences of vedolizumab and vedolizumab are Patent Publication No. 2014 / 0341885 and U.S. Patent Publication No. 2014 / 0377251 (the entire contents of each of which are expressly incorporated herein by reference in their entirety) α4β7 antibodies and their corresponding amino acid sequences are also described in U.S. Pat. 3752, which is incorporated herein by reference.

[0111] The use of the methods disclosed herein with other antibodies is also encompassed by the present invention. The methods described herein may be used to treat a variety of diseases, including but not limited to, inflammatory bowel diseases, including alpha-4 integrin-binding antibodies, including natalizumab. Alternatively, the method may be performed using an antibody or antigen-binding fragment thereof. The methods described herein include, but are not limited to, those directed against both integrins α4β7 and αEβ7. Antibodies or antigen-binding fragments thereof that bind to β7 integrins, including etrolizumab. fragments (the sequence of etrolizumab is 6833 (herein incorporated by reference).

[0112] IL-23 inhibitors As noted above, the methods disclosed herein can be used to treat autoimmune diseases, e.g., arthritis. inflammation or psoriasis, and / or inflammatory diseases, such as IBD (e.g., Crohn's disease or and administering to a subject with ulcerative colitis (e.g., ulcerative colitis) an α4β7 inhibitor, e.g., an anti-α4β7 antibody, or and an IL-23 inhibitor, e.g., an anti-IL-23 antibody or an anti-IL-23R. This involves administering both the antibody and the IgG antibody.

[0113] As used herein, the term "human interleukin-23" or "hIL-23" The term refers to the IL12B subunit (RefSeq accession number NM_002187 .3;NP_002178.2) and IL23A subunit (RefSeq accession Heterodimers formed by the fusion of NP_057668.1 and NM_016584.3 "IL12B" refers to a multimeric pro-inflammatory cytokine. Also known as FM2 and NKSF2; this p40 subunit mediates IL-12 expression. The amino acid sequence of p40 is set forth in SEQ ID NO:13. "IL23A" refers to p19, IL-23p19, IL-23 subunit α, and SGR Also known as F. The amino acid sequence of p19 is set forth in SEQ ID NO: 14. IL-2 3 binds to the receptor IL-23R, which is composed of two subunits, IL23R (R efSeq accession numbers NM_144701.3; NP_653302.2) and IL-12RB1 (RefSeq accession number NM_005535.3; NP_0 05526.1). Both subunits are required for IL-23 signaling. It is said that.

[0114] In certain embodiments, IL-23 inhibitors that can be used in the combination therapies disclosed herein are The inhibitors are the p19 subunit, p40 subunit, or their shared enzymes of human IL-23. Anti-IL-23 antibodies or antigen-binding fragments thereof that specifically bind to any of the above-mentioned IL-23 antigens. In certain embodiments, the compounds that can be used in the combination therapies disclosed herein are The IL-23 inhibitor is an anti-IL-23R antibody that specifically binds to human IL-23R or and an antigen-binding fragment thereof.

[0115] In one embodiment, an anti-IL-23 antibody that can be used in the combination therapy disclosed herein Brazikumab is a steroid drug that is used in the treatment of rheumatoid arthritis. Brazikumab is also known as MEDI-2070. Brazikumab is disclosed, for example, in U.S. Pat. 22033 and US9487580 (regarding brazikumab (including its sequence) (which is incorporated by reference in its entirety).

[0116] In some embodiments, anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein The antibody (e.g., anti-p19 antibody) is guselkumab. Guselkumab is also available as TREMFYA. (trademark) or CNTO1959. Guselkumab is disclosed, for example, in U.S. Pat. Patent Nos. US7491391 and US10030070 (guselkumab (including its sequence) ) which are incorporated by reference in their entirety. be.

[0117] In some embodiments, anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein The antibody (e.g., anti-p19 antibody) is risankizumab. Risankizumab is a compound of SKYRI Also known as ZI™, BI655066, or ABBV-066. Kizumab is disclosed, for example, in U.S. Pat. Nos. US8778346 and US9441036 ( Risankizumab (including its sequence, which is incorporated by reference in its entirety) These methods are known in the art, including those

[0118] In some embodiments, anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein The antibody (e.g., anti-p40 antibody) is briakinumab. Briakinumab is a steroid drug. 74, J695, BSF 415977, LU 415977, A-796874.0, and WAY-165772. Briakinumab is disclosed, for example, in US 691 4128, US7504485, US9072725, and US20150037348 (incorporated by reference in its entirety with respect to briakinumab, including its sequence) can be.

[0119] In some embodiments, anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein The antibody (e.g., anti-p19 antibody) is tildrakizumab. Tildrakizumab is administered by ILU. MYA™, tildrakizumab-asmn, MK-3222, SCH 900222 Tildrakizumab is also known as hum13B8-b, or hum13B8-b. Patent Nos. US8362212, US8404813, and US9803010 (incorporated by reference in its entirety with respect to tildrakizumab, including its sequence) These methods are known in the art, including those described herein.

[0120] In some embodiments, the anti-IL- 23 antibody (e.g., anti-p40 antibody) is ustekinumab. Ustekinumab is a CN Also known as TO1275, TT-20, C01275, or STELARA™ Ustekinumab is a compound described in the art, for example, in US6902734, WO2002 012500, US7279157, US20200095315 (ustekinumab (so The sequences of which are incorporated by reference in their entirety.

[0121] Another example of an anti-IL-23 antibody that can be used in the combination therapy disclosed herein is LY LY2525623 is disclosed, for example, in U.S. Pat. No. 7,872,102 No. and US9023358 (LY2525623 (including its sequence) in its entirety) (which is incorporated by reference) are well known in the art.

[0122] In one embodiment, an anti-IL-23 antibody that can be used in the combination therapy disclosed herein CNTO4088 is disclosed in, for example, U.S. Patent No. US7807 414 and US 7,935,344 (incorporated in their entirety by reference to CNTO 4088) These methods are well known in the art, including those described in

[0123] In some embodiments, the anti-IL- The antibody is mirikizumab, also known as LY3074828. Mirikizumab has been reported, for example, by Ma et al. (Expert Opinion Research Estig Drugs, 27:649-660 (2018)) and Reich et al. al.(Br J Dermatol,181:88-95(2019))(Milisaki (which is incorporated herein by reference in its entirety for mabs).

[0124] Other anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein include, for example, Publication Nos. US20110206686 and US20120264917, Nos. US9127057 and US7510709, and Li et al. (Anal Chem 89:2250-2258(2017)) (IL-23 antibody These methods are known in the art, including those described in the above, which are incorporated by reference in their entirety. Other anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein (e.g., anti-p19 Antibodies) are described, for example, in WO2007147019, WO2008134659, WO200 9082624, US8333968, US20110159589, US201202 94862, US20130309235, US9464134, and US201501 97566 (incorporated by reference in its entirety regarding IL-23 antibodies).

[0125] Other anti-IL-23 antibodies suitable for use in the combination therapies disclosed herein (e.g., anti-p 40 antibodies) are described, for example, in US20150010544, US9708395, and US2 No. 0160333085 (incorporated by reference in its entirety regarding IL-23 antibodies) will be done.

[0126] In certain embodiments, an IL-23 inhibitor suitable for use in the methods described herein is In some embodiments, the small molecule inhibitor of IL-23 is For example, by reducing or suppressing IL-23 production at the transcriptional or translational level, In some embodiments, small molecule inhibitors of IL-23 may reduce or inhibit the production of IL-23. For example, by reducing or inhibiting the interaction of IL-23 with the receptor (IL-23R), and / or reducing or inhibiting the ability of the subunits p19 and p40 to form a complex. By inhibiting IL-23, the function of IL-23 can be reduced or suppressed.

[0127] In some embodiments, small amounts of IL-23 suitable for use in the combination therapies disclosed herein are The molecular inhibitor is STA-5326, which prevents the nuclear translocation of c-Rel. STA-5326 inhibits the production of IL-23 by inhibiting the Also known as mesylate, Apilimod, and LAM-002. The chemical name of 5326 is (E)-4-(6-(2-(3-methylbenzylidene)hydrazinium) -2-(2-(pyridin-2-yl)ethoxy)pyrimidin-4-yl)morpholine STA-5326 is a dimethanesulfonate. al.(Inflamm Bowel Dis,12:558-565(2006)) and and Wada et al.(Blood,109:1156-1164(2007))( (which is incorporated by reference in its entirety).

[0128] In some embodiments, anti-IL-2 antibodies suitable for use in the combination therapies disclosed herein The 3R antibody is AS2762900-00. AS2762900-00 is, for example, US9556276, US20140275490, WO2012137676, US9 371391, US20150126713, WO2013129454, Imamur a et al. (Eur J Pharmacol, 824:163-169(201 8)), Sasaki-Iwaoka et al. (Eur J Pharmacol , 828:89-96(2018)), and Sasaki-Iwaoka et al. (Eur J Pharmacol, 843:190-198(2019))(IL-2 3R antibodies) which is incorporated by reference in its entirety.

[0129] Other examples of anti-IL-23R antibodies suitable for use in the combination therapies disclosed herein include, for example, US20100166767, WO2008106134, US201101589 92, WO2010027767, US20120148582, US8691532, and US20140170154 (incorporated by reference in their entirety regarding IL-23R antibodies). (The following is a list of the items to be included in the calculation):

[0130] Patient selection In some embodiments, patients are administered a combination of the drugs disclosed herein based on specific characteristics. In some embodiments, the patient is selected for treatment with an α4β7 inhibitory therapy. A non-responder to drug therapy. ", or "vedolizumab non-responders," are those who have not received α4β7 inhibitors, e.g., anti-α4β7 antibodies (e.g., have received the first and second induction doses of a therapy, e.g., vedolizumab, and Signs of non-response early in Ibuprofen treatment (e.g., approximately 3 or 4 weeks after the second induction dose) have an autoimmune disease and / or IBD (e.g., Crohn's disease or ulcerative colitis) Signs of non-response may include, for example, measures of clinical response and / or may include failure to achieve the measurements described herein. , the patient is a non-responder to α4β7 inhibitor therapy. "Non-remitters," "remitters," or "vedolizumab non-remitters" are defined as those with autoimmune and / or is a subset of patients with IBD (e.g., Crohn's disease or ulcerative colitis), The patient is treated with an α4β7 inhibitor, e.g., an anti-α4β7 antibody (e.g., vedolizumab), for the first and second time. Those who have received a second or third induction dose and are currently receiving therapy, e.g., vedolizumab therapy Signs of non-remission early in the course (e.g., approximately 3 or 4 weeks after the second or third induction dose) For example, patients may show a response rate of 0 and 2 weeks to an anti-α4β7 antibody (e.g., vedolizumab). Patients may receive treatment at 0, 2, and 6 weeks. However, for example, about 3 to 4 weeks after the second administration, the efficacy of, for example, an anti-α4β7 antibody (e.g., 5 or 6 weeks after the start of treatment with vedolizumab, or approximately 3 to 8 weeks after the third dose Clinical remission may not be achieved after weeks, for example, 10 or 14 weeks after initiation. Indications of resolution may include, for example, measurements of clinical remission and / or achieving the measurements described herein. This may include not being able to do so.

[0131] In some embodiments, the non-response is due to, but not limited to, an α4β7 inhibitor, e.g., an anti- Using an algorithm that includes components including α4β7 antibody concentration and / or antibody clearance Antibody concentrations may be measured in serum obtained from the patient. In some embodiments, elements of the algorithm for identifying non-effective treatments include weight and / or Non-responders to α4β7 inhibitor therapy may include, for example, high blood pressure, urinary tract infection, or albumin levels. Low inhibitor clearance, low albumin levels, high fecal calprotectin levels, early There is no decrease in fecal calprotectin levels after treatment and / or patients may have a higher body weight. stomach.

[0132] In some embodiments, the patient has a high level of a marker gene, e.g., IL-22, STAT5 The subject is characterized by elevated levels of IL-1β and / or IL-1β. In this condition, the levels of IL-22, STAT5A, and / or IL-1β were 5%, 10%, , 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% , 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or or, in certain embodiments, IL-22, STAT5A, and / or The IL-1β levels may be measured, for example, in a control sample from a non-IBD subject, e.g., a healthy human, or or compared to control levels in control non-inflamed colon tissue or non-colon tissue obtained from patients. 5% to 35% compared with the reference level for responders to α4β7 inhibitors, 5%~25%, 5%~20%, 5%~15%, 5%~10%, 10%~20%, 10%~ 15%, 10%~30%, 15%~40%, 20%~50%, 25%~60%, 30%~ 70%, or 40% to 100% or more. Elevated levels are The biopsy sample may be taken before treatment (e.g., 2 to 3 days after treatment). 10 days prior), or an α4β7 inhibitor, such as an anti-α4β7 antibody, such as vedolizumab, Measurements can be obtained the day before or on the first day of treatment with at the nucleic acid level, or by, for example, immunohistochemistry using methods well known in the art. The levels of proteins involved may also be tested.

[0133] In some embodiments, IL-22 (interleukin-22) is obtained from National l Center for Biotechnology Information At least 95% of SEQ ID NO: 9 in the database maintained by the University of Leland, Bethesda, , 97%, 99%, or 100% identical to gene ID 50616 (GenBank Accession number NM_020525.5), or the protein of SEQ ID NO: 10 (Ge nPept has the accession number NP_065386.1. In some embodiments, STAT5A (signal transducer and activator of transcription 5A) is ional Center for Biotechnology Informati SEQ ID NO: 11 and a few others in the database maintained by the NIH Pharmacy, Inc. (Bethesda, MD) Gene ID 6776 (Ge nBank accession number NM_003152.3), or the protein of SEQ ID NO: 12 Protein (represented as GenPept accession number NP_003143.2), In some embodiments, IL-1β (interleukin-1β) is ional Center for Biotechnology Informati GenBank accession numbers in the database maintained by the National Institute of Standards and Technology (NIS) in Bethesda, Maryland. Session number NM_000576 and at least 95%, 97%, 99%, or 100% % identical to gene ID 3553, or GenPept accession number NP_00 0567.

[0134] In some embodiments, the patient to be administered the combination therapy is an α4β7 inhibitor, e.g. , at the start of treatment with an anti-α4β7 antibody, e.g., vedolizumab, or at 6 or 10 weeks after the start of treatment In some embodiments, the patient is characterized by elevated levels of IL-22 in the serum. , patients to be administered the combination therapy may be receiving an α4β7 inhibitor, e.g., an anti-α4β7 antibody, e.g., , elevated serum IL-22 levels at the start of or 6 weeks after initiation of vedolizumab treatment This is characterized as occurring between the onset and 10 weeks, between the onset and 6 weeks, or between 6 weeks and 10 weeks. No reduction, less than half reduction, or less than one-third reduction in the first week In some embodiments, the elevated level of IL-22 in serum is greater than 3 pg / ml, 5 pg / ml, or ~100pg / ml, >5pg / ml, >10pg / ml, 10~120pg / ml, 1 In some embodiments, in remission subjects, blood levels are greater than 0 pg / ml or higher. The levels of IL-22 in the supernatant were less than 10 pg / ml, less than 5 pg / ml, and less than 3 pg / ml. or decrease to less than 2.7 pg / ml, less than 2 pg / ml, or after initial treatment, e.g. In some embodiments, the non- In patients in remission, serum IL-22 levels were below 10 pg / ml and below 5 pg / ml. or does not decrease to less than 3 pg / ml, less than 2.7 pg / ml, or less than 2 pg / ml, or is not undetectable after initial treatment, e.g., by week 6 and / or 10 of treatment. stomach.

[0135] In some embodiments, the patient to be administered the combination therapy is an α4β7 inhibitor, e.g. , at the start of treatment with an anti-α4β7 antibody, e.g., vedolizumab, or at 6 or 10 weeks after the start of treatment In some embodiments, the patient is characterized by elevated levels of fecal calprotectin. In certain embodiments, the patient to be administered the combination therapy may be an α4β7 inhibitor, e.g., an anti-α4β7 antibody, For example, fecal calprotectin levels at the start of or 6 weeks after initiation of vedolizumab treatment characterized by an increase in the level of BP, which may be from onset to 10 weeks, from onset to 6 weeks, or Between weeks 6 and 10, there was no reduction, a reduction of less than half, or a reduction of less than one-third. See, e.g., US2017360926, which is incorporated by reference in its entirety. In some embodiments, the patient to be administered the combination therapy has a high level of fecal calprostaglandin. Compared to normal levels of tektin, 5%, 10%, 15%, 20%, 25%, 30%, 3 5%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 8 5%, 90%, 95%, 100% or more, or 5% to 35%, 5% to 25 %, 5%~20%, 5%~15%, 5%~10%, 10%~20%, 10%~15%, 1 0%~30%, 15%~40%, 20%~50%, 25%~60%, 30%~70%, 4 0% to 100% or greater increase in fecal calprotectin levels It can be characterized as being.

[0136] Compared to normal levels, or in certain embodiments, IL-22, STAT5A, and and / or IL-1β levels are elevated. In some embodiments, the combination therapy is administered Patients who should be treated with an α4β7 inhibitor, e.g., an anti-α4β7 antibody, e.g., vedolizumab, should be considered for initiation of treatment. Both IL-22 and fecal calprotectin at baseline, 6 weeks, and 10 weeks after initiation In some embodiments, the patient is administered a combination therapy and characterized by elevated levels of Patients who should be treated with an α4β7 inhibitor, e.g., an anti-α4β7 antibody, e.g., vedolizumab Levels of both IL-22 and fecal calprotectin at baseline or 6 weeks after initiation This is characterized by an increase in the serotonin level between the start and 10 weeks, between the start and 6 weeks, or between the start and 6 weeks. Between the 1st and 10th week, there was no reduction, a reduction of less than half, or a reduction of less than one-third. Decreases.

[0137] In some embodiments, the patient to be administered the combination therapy is a non-responder, e.g. , clinical response to vedolizumab at 6 and / or 10 weeks after initiation of vedolizumab treatment In some embodiments, the patient to be administered the combination therapy is a non-remission For example, vedolizumab at week 6 and / or week 10 after initiation of vedolizumab treatment No clinical remission has been observed.

[0138] As used herein, the term "sample" refers to a sample, e.g., a tissue, isolated from a subject. Tissues, cells, biological fluids, and isolates thereof, as well as tissues, cells, biological fluids, and isolates thereof present within a subject and present in a patient or is intended to include tissues, cells, and fluids that can be obtained from normal subjects. For example, non-invasive methods for in vitro measurement of markers that distinguish responders or non-responders are available. The experimental sample may include a blood or serum sample. Thus, the blood sample may be analyzed for marker characteristics, e.g., For example, testing for size, sequence, composition, activity, or quantity (i.e., level) For patients with autoimmune diseases or IBD, normal marker characteristics, e.g. A control, reference sample, for size, sequence, composition, activity, or quantity (i.e., level) of IB It can be obtained from a healthy subject who does not have D.

[0139] The blood collection container may contain an anticoagulant, e.g., an additive that preserves the integrity of the blood, e.g., dextromethorphan. Heparin or ethyl acetate, containing phosphate or albumin or buffer, e.g., phosphate May contain diaminetetraacetic acid (EDTA), sodium citrate, or citric acid solution If the amount of the marker is measured by measuring the level of DNA in the sample, DNA stabilizers, such as agents that inhibit DNAses, can be added to the sample. If the amount of a target molecule is measured by measuring the level of RNA in a sample, R NA stabilizers, such as agents that inhibit RNAses, can be added to the sample. If the amount of carcinoma is measured by measuring the level of protein in a sample, Protein stabilizers, e.g., agents that inhibit proteases, can be added to the sample. An example of a blood collection container is a PAXGENE™ tube ( PREANALYTIX, Valencia, CA). Peripheral blood samples were collected using modified For example, it can be fractionated, sorted, or enriched (eg, in the case of a reference sample).

[0140] The sample, e.g., blood or modified blood, and / or a reference, e.g., a matched control (e.g., , germline), the sample is analyzed for marker genes (e.g., IL-22, STAT5A, and / or IL-1β), such as size, sequence, composition, activity, or quantity (i.e., Before assessing the level of and / or protein extraction, fixation, storage, freezing, ultrafiltration, concentration, evaporation, centrifugation, etc. ) can be applied.

[0141] In some embodiments, the marker is a nucleic acid marker, e.g., DNA, RNA, cDNA, or NA, or marker genes, such as IL-22, STAT5A, and / or IL- Protein markers that correlate with 1β can be identified by sequencing. There are several sequencing methods known in the art for sequencing nucleic acids. The primer or primer pair is a primer that is used to identify one or more DNA fragments corresponding to the marker gene. can be used to sequence both strands. The region of interest was identified before sequencing to increase the amount of sequence for gene detection. To amplify, a probe, e.g., a nucleic acid probe, e.g., a hybridization probe, It can be used in combination with a probe. Examples include whole genes, transcripts of genes, and fragments or transcripts of genes, e.g. For example, one or more exons or untranslated regions, or a portion of the marker containing the mutation site, Examples of target sequences for primer selection and sequence or composition analysis include: collects sequence or mutation information, e.g., RefSeq, COSMIC, and dbGaP These can be found in public databases that collect

[0142] Sequencing methods are known to those skilled in the art. Exemplary methods include the Sanger method, SE These include the QUENOM™ method and next-generation sequencing (NGS) methods. To separate the extended labeled DNA fragments, electrophoresis, e.g., capillary electrophoresis, is used. The Sanger method, which involves the use of electrophoresis, can be automated for high-throughput applications. Primer extension sequencing is performed after PCR amplification of the region of interest. It is possible.

[0143] In some embodiments, DNA markers, e.g., marker genes (e.g., IL- Genomic DNA of the IL-1β gene (e.g., STAT5A, STAT6A, and / or IL-1β) is available from known methods in the art. in biological samples using the method, in situ and in vitro formats DNA can be analyzed both directly from the sample and as a separate cellular component. The kit, e.g., RNA or protein, can be isolated after isolation. For example, the QIAAMP™ DNA Micro Kit (Qiagen, CA) Valencia) are available for DNA isolation. DNA can be isolated using such kits. It can also be amplified.

[0144] In another embodiment, marker genes (e.g., IL-22, STAT5A, and / or mRNA markers for IL-1β can be identified using methods known in the art. Samples can be analyzed in both in situ and in vitro formats Many expression detection methods use isolated RNA. For in vitro methods, mRNA Any RNA isolation technique that does not select for the isolation of RNA from cells is suitable. (e.g., Ausubel et al., ed., Curren t Protocols in Molecular Biology,John Wi ley & Sons, New York 1987-1999). In addition, many organizations Samples were prepared using, for example, the single-step RNA isolation process of Chomczynski ( 89, U.S. Pat. No. 4,843,155, or the like. RNA can be processed according to standard procedures (e.g., Chomczynski et al., 2004). and Sacchi (1987) Anal.Biochem.162:156-159 ), solutions (e.g., trizol, TRI REAGENT™ (Molecula r Research Center, Inc., Cincinnati, Ohio; U.S. Patent No. No. 5,346,994) or kits (e.g., QIAGEN™ Group RNEASY™ Isolation Kit (Valencia, CA) or LEUKO LOCK™ Total RNA Isolation System, Ambion division of Applied Biosystems , Austin, TX).

[0145] An additional step of removing DNA from the RNA sample may be used. Ionic detergent, followed by microcentrifugation to remove nuclei and therefore most of the cellular DNA DNA can then be isolated from the nuclei for DNA analysis. In one embodiment, the RNA can be isolated by guanidine thiocyanate lysis followed by Using CsCl centrifugation to separate RNA from DNA, various types of cells of interest can be analyzed. Extracted from the vesicles (Chirgwin et al. (1979) Biochemist ry 18:5294-99). Poly(A) + RNA is purified using oligo dT cellulose. Selection by selection (Sambrook et al. (1989) Molecula r Cloning--A Laboratory Manual(2nd ed.), Cold Spring Harbor Laboratory,Cold Spring (See e.g., Harbour, NY). Alternatively, RNA can be separated from DNA using methods such as Organic extraction using hot phenol or phenol / chloroform / isoamyl alcohol This can be achieved by extraction. If necessary, an RNAse inhibitor can be added to the lysis buffer. Similarly, certain cell types may require a protein denaturation / digestion step. For many applications, it may be desirable to add transfer RNA to the mRNA in relation to other cellular RNAs such as tRNA (tRNA) and ribosomal RNA (rRNA) It is desirable to enrich for most mRNAs, as they contain poly(A) tails at their 3' ends. This can be achieved by affinity chromatography, e.g., on cellulose or SEPHA. Oligo(dT) or poly(U) bound to a solid support such as DEX™ media It is possible to enrich them using 4)Current Protocols In Molecular Biology ,vol.2,Current Protocols Publishing,New (See York). Upon binding, poly(A)+ mRNA is eluted in 2 mM EDTA / 0. Elution from the affinity column is performed using 1% SDS.

[0146] Analyzing the characteristics of the marker genes described herein in a biological sample can be used to identify the test subject. The characteristics include obtaining a biological sample (e.g., a blood sample or a reference sample) from For example, nucleic acids (e.g., RNA, mRNA, genomic DNA, or cDNA) and / or Detecting or detecting a characteristic of the translated protein, e.g., a marker or markers It can be assessed by any of a wide variety of well-known methods for measuring such. Non-limiting examples of suitable methods include methods for detecting secreted, cell surface, cytoplasmic, or nuclear proteins. immunological methods, protein purification methods, protein function or activity assays, optionally The matched regions, i.e., the regions of variability or diversity, are digested, and the resulting digested fragments are The method includes a "mismatch cleavage" step to separate and identify the variant or variants from the target gene (Myer S, et al. (1985) Science 230:1242) Nucleic acid hybridization These include nucleic acid amplification, nucleic acid reverse transcription, and nucleic acid amplification and analysis of the amplified products. These methods involve gene array / chip technology, e.g., RT - PCR, TAQMAN™ gene expression assay (Applied Biosystems ems, Foster City, CA), in situ hybridization , immunohistochemistry, immunoblotting, FISH (fluorescence in situ hybridization) ation), FACS analysis, Northern blot, Southern blot, INFINIUM (commercial Target) DNA analysis bead chip (Illumina, Inc., Sandy, CA) Ego), quantitative PCR, bacterial artificial chromosome array, single nucleotide polymorphism (SNP) array (Affym etrix, Santa Clara, CA) or cytogenetic analysis.

[0147] Detection methods include, for example, detecting RNA in biological samples in vitro and in vivo. A. Can be used to detect mRNA, protein, cDNA, or genomic DNA. Furthermore, the present invention can be applied to the detection of polypeptides or nucleic acid markers described herein. In vivo techniques involve the use of markers, e.g., markers or labeled antibodies, Fc receptors or The present invention provides a labeled protein that detects an antigen to a polypeptide, e.g., a wild-type or mutant marker. For example, antibodies can be labeled with a radioisotope. Its presence and location within the subject can be detected by standard imaging techniques. These assays can be performed in a variety of ways. Those skilled in the art will readily identify the marker(s) of interest. (Acceptable), these or other appropriate and available methods based on the tissue sample and the nature of the mutation. You can choose from several methods, some of which are explained in more detail in later sections. The method or combination of methods may be used in different cases or, for example, in different patients. In populations, this may be appropriate.

[0148] In vitro techniques for detecting polypeptides corresponding to a marker of the invention include: Enzyme-linked immunosorbent assay (ELISA), Western blot, protein array, immunoassay In such instances, the expression of the marker can be measured by antibody ( (e.g., unlabeled, radiolabeled, chromophore-labeled, fluorophore-labeled, or enzyme-labeled antibodies) , antibody derivatives (e.g., proteins in substrates or protein-ligand pairs (e.g., or antibodies conjugated to a ligand (e.g., biotin-streptavidin )), or a marker protein that undergoes all or some of the normal post-translational modifications, Marker proteins or fragments thereof, e.g., containing regions that can be mutated Proteins or fragments containing mutated residues or mutated sequences or structural contexts antibody fragments (e.g., single chain antibodies, isolated antibody hypervariable domains) that specifically bind to the The antibody is selected from the group consisting of SEQ ID NOs: 10 and 12. A sandwich ELISA assay can detect proteins having an amino acid sequence corresponding to the target protein. Any assay may, for example, measure the gain or loss of the amount of a marker in a sample compared to a reference sample. A standard ELISA can detect the loss of antibody binding between patient and reference samples. The levels will be compared.

[0149] In some embodiments, the method involves measuring the amount (i.e., level) of a marker protein. In some embodiments, the amount of a marker protein is determined, for example, by measuring Ve Antana Medical Systems (Arizona) or Illumina (San Diego) Quantification is performed by immunohistochemistry of the samples using the quantification products and methods of San Diego. In some embodiments, the amount of marker protein is determined by immunohistochemistry of the marker from blood. In some embodiments, the amount of marker protein is quantified by antibody binding or In some embodiments, the marker protein is determined by a score of staining intensity. The amount of isoform in healthy cells or tissue versus cells from a subject with an autoimmune disease or IBD. or by comparison of antibody binding or staining in tissue.

[0150] In one embodiment, marker expression is measured by extracting mRNA / cDNA (i.e., by preparing a marker nucleic acid or a fragment thereof. Hybridize the mRNA / cDNA with a reference polynucleotide that is the complement of the fragment The cDNA is optionally hybridized with a reference polynucleotide. amplification using one of a variety of polymerase chain reaction methods prior to depolymerization Similarly, the expression of one or more markers can be measured by measuring the expression level of the marker(s). Quantitative PCR can be used to assess mRNA levels. For example, an inactivating mutation in a marker gene can result in changes in mRNA levels within the cell. The levels are a feedback loop that takes into account non-functional or non-existent proteins. Upregulation of signaling proteins for protein production or instability of modified mRNA sequences Alternatively, a mutant or variant of a marker of the invention may be downregulated due to its phenotype. Any of the many known methods for detecting mutations (e.g., the single nucleotide polymorphisms, deletions, etc. mentioned above) can be used to identify the patient. The method may be used to detect the occurrence of mutations in marker genes in a subject.

[0151] An example of a direct measurement is the quantification of a transcript. As used herein, the level of expression or The amount may be the absolute amount of expression of the mRNA encoded by the marker, or the amount of expression of the mRNA encoded by the marker. It refers to the absolute expression level of the encoded protein. It is based on the absolute expression level of the selected marker. Instead of making the determination based on the expression level, the determination may be based on a normalized expression level. For example, in a constitutively expressed housekeeping role, its expression is not a marker. By correcting the absolute expression level of the marker when compared to the expression of a control marker that does not contain the marker. Suitable markers for normalization include the actin gene or β Housekeeping genes such as 2-microglobulin are also included. Reference markers for this purpose include those that are ubiquitously expressed and / or whose expression is associated with oncogenes or Constitutively expressed genes include markers that are not regulated by cytokines. , according to the relevant tissue and / or patient context and analytical methods known in the art. Such normalization can be performed, for example, at different times or at different It is possible to compare expression levels in one sample with another sample between subject samples. The expression level can be provided as a relative expression level. Initially, for example, the diploid copy number is determined by comparing cells derived from a subject who does not have an autoimmune disorder or IBD. The correlation of a marker or marker set can be determined by measuring the amount of To determine the relative abundance, the amount of a marker or marker set is determined by the expression level for the sample in question. Before determining your current level, you should use at least 1, 2, 3, or 4 to determine your starting point. , 5, or more samples, e.g., 7, 10, 15, 20, or 50, or The assay is performed on a number of samples to establish the starting measurement. The average amount or level of each of the selected markers or marker sets is determined, is the starting expression level for the biomarker or set of biomarkers in question. The amount of the marker or set of markers determined for the test sample ( The absolute level of expression (e.g., expression level) is used to determine the expression profile obtained for that marker or marker set. Divide by the starting value. This provides a relative amount and indicates abnormal levels of marker protein activity. It helps to identify.

[0152] Probes based on the sequences of the nucleic acid molecules of the invention correspond to one or more markers of the invention. It can be used to detect transcripts or genomic sequences. The label may include an attached label group, e.g., a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes may, for example, encode proteins in a sample of cells from a subject. For example, by detecting mRNA levels or by measuring the levels of nucleic acid molecules that correspond to the target gene. by determining whether genes encoding the gene or protein are mutated or deleted. Use as part of a diagnostic test kit to identify cells or tissues that express the protein It is possible.

[0153] The primer or nucleic acid probe contains a nucleotide sequence complementary to a specific marker. selectively hybridize to the marker gene or nucleic acid associated with the marker gene. The primers and probes are of sufficient length to allow for the isolation and sequencing of the marker nucleic acid. In one embodiment, a primer or nucleic acid sequence can be used to aid in sequencing. The nucleic acid probe, e.g., a substantially purified oligonucleotide, is about 600 nucleotides from the marker gene. , 8, 10, 12, or 15, 20, 25, 30, 40, 50, 60, 75, 100 or more consecutive nucleotides (within ±5% of the value) that hybridize under stringent conditions. In another embodiment, the primer or nucleic acid The probe may comprise the nucleotide sequence of any of the sequences set forth in either SEQ ID NO: 9 or 11. The marker nucleic acid can hybridize to a sequence comprising at least about 1 comprises a nucleotide sequence of 5 consecutive nucleotides, at least about 25 nucleotides, or The primer has about 15 to about 20 nucleotides set forth in either SEQ ID NO: 9 or 11. primer or nucleic acid probe. Nucleic acid analogs serve as binding sites for hybridization. An example of a suitable nucleic acid analog is a peptide nucleic acid (e.g., E Gholm et al., Nature 363:566 568(1993); USA See Patent No. 5,539,083).

[0154] The primer or nucleic acid probe is characterized by binding energy, base composition, sequence complexity, cross-linking, and It uses an algorithm that takes into account hybridization binding energy and secondary structure. (Friend et al., International Journal of Clinical Chemistry, 2019, 133:111–112, 2019). l Patent Publication WO01 / 05935,publishe d Jan.25,2001;Hughes et al., Nat.Biotech. 19:342-7 (2001). Those skilled in the art will appreciate that, using techniques in the art, e.g., By manipulating the degeneracy or GC content in the dimer or nucleic acid probe, The possibility of a primer or nucleic acid probe that binds to a sequence, mutant, or allelic variant The ability to detect markers or similar characteristics disclosed herein can be used to Related markers, e.g., markers on chromosomal loci or the same markers described herein Primers and nucleic acid probes can be designed for mutations in different regions of the CAR gene. A computer program well known in the art is Oligo version 5.0 (Na (National Biosciences, Plymouth, Minnesota) It is useful for designing primers with optimal amplification properties and isotopic identity. The probes and primers may be used to identify the markers and their variants, polymorphisms, or alleles described herein. can be used to detect genes, and deviations from perfect complementarity are This is contemplated if the removal does not prevent the molecule from specifically hybridizing to the target region. For example, an oligonucleotide primer may have a non-complementary fragment at its 5' end. The remainder of the primer may have a non-complementary nucleotide. The nucleotide sequence is used to ensure that the resulting probe or primer still specifically hybridizes to the target region. The nucleic acid probes or primers may be interspersed with the nucleic acid probes or primers as long as they are capable of hybridizing with each other.

[0155] The following examples illustrate the improvements to the methods and compositions described herein. It is provided for illustrative purposes only and is not intended to limit the scope of the invention in any way. Commercially available reagents referred to in the examples were used according to manufacturer's instructions unless otherwise indicated. was used. [Example]

[0156] Example 1. Combination of anti-MAdCAM-1 antibody and anti-p40 antibody in a mouse colitis model Effectiveness of the combination The following studies have demonstrated the efficacy of anti-MAdCAM-1 antibodies and IL-23 inhibitors, particularly in the treatment of murine colitis models. We used IL-23 (and IL-12) to bind to and block the p40 subunit of IL-23 (and IL-12) To evaluate the combined therapeutic effect of a combination therapy containing two therapeutic agents, as described below. Anti-MAdCAM-1 (MECA-367 from BioXCell (catalog number BE00 35)) and anti-p40 IL-23 / IL-12 (clone C17 from BioXCell) The combination of .8 (Cat. No. BE0051) significantly reduced colon weight in tested mice vs. controls. This resulted in improvements in mass, diarrhea, and histopathology.

[0157] The model used was the naive T cell transfer (TCT) model. Infectious colitis is primarily driven by a Th1 / Th17-mediated immune response. The disease is characterized by infiltration of the lamina propria with CD4+ T cells, neutrophils, and macrophages. This leads to progressive weight loss, inflammation of the colon, and diarrhea.

[0158] A schematic diagram of the study is shown in Figure 3. In summary, naive T cells ( CD4+CD62L+ SCID mice (approximately 2 x 10 5 cells / mouse). sotypic control antibody ("vehicle"), 10 mg / kg anti-MAdCAM antibody, 1 mg / kg g of anti-p40 IL12 / IL23 antibody (anti-p40 antibody), or 10 mg / kg of anti-M The combination of AdCAM antibody and 1 mg / kg anti-p40 IL12 / IL23 antibody Mice were treated.

[0159] As shown in Figure 3, vehicle or 10 mg / kg anti-MAdCAM antibody was administered on day 0. The subjects were administered the following doses on days 7, 14, 21, and 28 (Q1W). 1 mg / kg anti-p40 IL12 / IL23 antibody every 4 days from day 14 to day 26 Analyses, e.g., diarrhea score, colon weight, colon histology, colon m Samples were collected for RNA.

[0160] The results of the study are provided in Figures 4A, 4B, 4C, Figures 5A and 5B, and Figures 6A and 6B. will be done.

[0161] The combination of anti-MAdCAM antibody and anti-p40 IL12 / IL23 antibody was administered on day 28. showed a significant effect on colon weight as shown in the results provided in Figure 4A. The benefit was also observed in the diarrhea scores shown in Figure 4B on day 21, but the diarrhea scores were significantly lower than those in Figure 4 A positive trend was observed for the data shown in Figure 4B on day 28 (data not shown) versus day 21. C provides a graph of histopathology scores at day 28 of H&E staining, showing the effect of antibody treatment alone or The combination therapy showed superior therapeutic efficacy compared with the control.

[0162] Furthermore, CD3 (T cell) staining of the lamina propria / epithelium on day 28 was significantly higher than that of the anti-MAdCAM1 antibody. As shown in Figure 5A, the primary effect was on T cell infiltration. Skin T cells were quantified for each treatment group. As described in Figure 5B, T cell infiltration was measured by total It correlated with the inflammatory cell score.

[0163] Anti-p40 antibody reduces the proportion of MPO(+) neutrophils in the colonic mucosa and anti-MAdCAM1 The combination of anti-p40 IL-23 / IL-12 inhibitors and anti-p40 IL-23 / IL-12 inhibitors was shown to be effective in macrophages (C The myeloperoxidase activity of neutrophils in the lamina propria on day 28 (D68) tended to decrease. MPO staining showed that the methyltransferase (MPO) staining was significantly increased in the IL-14 / IL-2 cells compared with the control cells. These results are presented in Figures 6A and 6B.

[0164] The results of this study showed that inhibition of the MAdCAM-1 pathway and the IL-23 pathway resulted in These findings suggest that they have complementary effects on adaptive and innate immune cells in the intestinal mucosa. Specifically, anti-MAdCAM-1 and anti-p40 inhibited the proliferation of CD3(+) T cells in the colonic mucosa. The anti-p40 antibody activity was also observed in the colon. The percentage of mucosal MPO(+) neutrophils was reduced, but CD68(+) macrophages were increased. was not significantly affected by MAdCAM-1, anti-p40, or their combination. (However, a trend was observed in the latter case.)

[0165] Example 2. Anti-MAd mRNA expression in a TCT-induced mouse chronic colitis model Combination effect of CAM-1 and anti-p40 We investigated gene expression in the colon and used anti-MAdCAM-1, anti-p40 IL-23, etc. Additionally, the effects of administering a combination of MAdCAM-1 and p40 IL-23 were determined. A false discovery rate (FDR) of 0.05 was used to define differentially expressed genes (DEGs). Figure 7 provides a Venn diagram showing gene expression counts. When the two treatments are combined, 4 ,422 genes were altered in expression, and approximately 3489 genes were synergistically expressed. The results showed that the differential gene expression between treatment and control is depicted in the graph in Figure 7. can be.

[0166] The combination of anti-MAdCAM1 antibody and anti-p40 antibody resulted in synergistic expression (upregulation or This resulted in a large number of genes showing upregulation (up or down-regulation) by anti-MAdCAM-1 and anti-p Genes that are affected only by the combination of 40 but not by individual treatments These synergistically expressed genes are Itgal (αL chain / CD11a / L FA-1A), Itgb2 (β2 integrin chain / CD18), Itgax (αX chain / C D11c), specific integrin chains such as Itga3, Itga9, and Itgb1bk In particular, the mRNA expression of Itgal (αL chain / CD11a / LFA-1A) , Itgb2 (β2 integrin chain / CD18), Itgax (αX chain / CD11c), Itga3 and Itgb1bk were synergistically down-regulated. In contrast, Itga9 , were shown to be synergistically upregulated.

[0167] In addition, certain cytokine genes, such as Il21r, Il12rb1, Il12a, and I L2ra, IL10ra, Il17re, Il34, Il18rap, Il1rl1, I l1b, Il1r2, Il3ra, Il1f9, Il23a, Iltifb, Il6, I Synergistic mRNAs in combination therapy, including l18bp, Il1a, Il15, and Il1r1 In particular, Il21r, Il12rb1, Il12a, and IL2r a, IL10ra, Il34, Il18rap, Il1rl1, Il1b, Il1r2, Il3ra, Il1f9, Il23a, Iltifb, Il6, Il18bp, Il1a Il1r1 and Il1r2 were synergistically down-regulated. In contrast, Il17re and Il15 were down-regulated. , were synergistically upregulated. Other genes that were downregulated (vs. therapy alone) were Stat4, Stat2, and Cd3g (synergistically downregulated). The study included a method control.

[0168] Overall, anti-MAdCAM-1 antibody treatment significantly reduced the number of ITGB7, IT It induced a wide range of gene expression changes, including GAE and interleukin receptors. Antibody treatment also significantly reduced the number of inflammatory cytokines induced by anti-MAdCAM-1 when compared to controls. Anti-MAdCAM-1 antibody treatment and anti-p40 treatment induced distinct and widespread changes in gene expression. Combined antibody treatment inhibits the expression of interleukins, integrins, stats, and Cd3 Broad gene expression was induced for approximately 4422 genes, including increased downregulation. IL12 Synergistic inhibition of IL23A and complementary inhibition of ITGB7 and ITGA4 indicate that combination therapy demonstrate that the drug exerts its effect in the "right" direction on one target. Repression does not increase expression of the remaining targets.

[0169] Example 3. Combination of anti-MAdCAM-1 and anti-p40 antibodies in T cell transplantation (TCT) The activity induced a mouse chronic colitis model. The experiment described in Example 1 was repeated using the treatment groups summarized in Table 1. [Table 1] Samples were collected and analyzed for α4β7+ CD4+ T cells, diarrhea score, colon weight, colon histology, and and colon mRNA were analyzed.

[0170] The combinations were administered on days 21 and 28, as shown in Figures 8A and 8B, respectively. The rats showed an improvement in body weight and a significant improvement in diarrhea score on day 21 (Fig. 8C). Unlike the results described in Example 1, no changes in mouse colon weight or tissue were observed on day 28. No significant effect on tissue pathology was observed with combination therapy.

[0171] The effect of the combination therapy on T cell, neutrophil, and macrophage infiltration was observed, and in Example 1 The results of this replicate study are presented in Figures 9A-9C. MAdCAM-1 antibody treatment and anti-p40 antibody treatment significantly increased the proportion of CD3(+) T cells in the colonic mucosa. The combination of anti-p4 Antibody treatment reduced MPO(+) neutrophils in the colonic mucosa (as shown in Figure 9B), suggesting that CD 68(+) macrophages were significantly affected by anti-p40 antibody treatment and combination therapy (Illustrated in Figure 9C). Overall, the results are very similar to those described from the study in Example 1. were similar.

[0172] Additionally, RNAseq was used to determine gene expression in the colon, with an FDR of 0.05. Approximately 343 genes showed novel and synergistic gene expression in the combination therapy. It was confirmed that it has.

[0173] To summarize the studies described in Example 1 and above, significant differences in diarrhea scores on day 21 were observed. A positive effect was observed in both studies. A trend towards improved weight gain from days 21 to 28 with the combo. was shown in a repeat study (but not in Study 1), but colon weight and histopathology in the first study The superior combo effect on HIV was not replicated in a second study. The complementation of T cells, neutrophils, and macrophages in the colon is very similar across studies. Overall, the two studies demonstrated that the MAdCAM-1 pathway and IL-23 Observations suggesting that inhibiting both p40 subunits provides significant activity. Furthermore, RNA-seq results demonstrated synergistic gene expression by the combination therapy. showed changes in

[0174] Example 4. Differential Expression of the IL-22 Pathway in Vedolizumab Responders vs. Non-Responders to IBD Patients adjustment Vedolizumab (VDZ) is a treatment for moderate to severe Crohn's disease (CD) and ulcerative colitis (U). Monoclonal antibodies (mAbs) targeting the α4β7 integrin have been approved for the treatment of b) Due to its inherent safety profile, VDZ is not a biologic that increases efficacy. Vedolizumab may be an excellent choice as the backbone of a comprehensive combination therapy. Blocks the interaction between AM-1 and α4β7.

[0175] Anti-p19 (interleukin-23 (IL-2)) with an acceptable safety profile 3) The mAb is effective in CD and UC. However, it still lacks the anti-p19 and VD It does not indicate whether the Z combination has additive efficacy.

[0176] Serum IL-22 levels have been shown to be higher in CD patients who respond to anti-p19 therapy. This suggests that increased activity of the IL-22 pathway is associated with the success of this treatment. Therefore, IL-22 may be an anti-inflammatory agent that provides superior efficacy when increased in VDZ non-responders. It is hypothesized that it supports combination with p19.

[0177] The publicly available microarray data were originally collected from 41 patients with moderate / severe biomarkers. The data were generated in patients with UC. Analysis was performed using a robust multichip averaging method. Calculate the log2 expression values ​​of IL22 and STAT5A probes in the array data, V Compare DZ responders, non-responders, and healthy controls. Moderate in the Gemini-III trial / CD patients with severe disease; 59 remitters (VDZ-R) at weeks 6 and 10, and both In 59 non-remission patients (VDZ-NR) at the time of registration, Quantikine Serum IL-22 levels were measured using a ELISA (R&D Systems). The mean CDAI at baseline was 297±49; the median CRP was 9.7mg / L [3. 7-24.5]; median FCP 583 mg / kg [229-1380]; 74.5% Anti-TNF deficiency). CDAI and FCP were used to assess disease severity at the start of treatment. and non-remission subjects were matched.

[0178] Baseline colonic IL22 mRNA levels were significantly higher in VDZ non-responder UC patients than in healthy controls. (endoscopic subscore > 1), but STAT5A mRNA (IL-22 signaling ) was higher in VDZ non-responders compared with responders (Figure 1). In addition, serum I L-22 was associated with a significant improvement in the efficacy and safety of VDZ in remission (CDAI<150) and non-remission CD patients. Although the decrease in IL-22 levels at week 10 was smaller in VDZ non-remitters, (Figure 2).

[0179] Response to VDZ is predominant in IBD patients in the absence of increased IL-22 pathway activity This suggests that the addition of anti-p19 / IL-23 therapy is complementary to VDZ and These results suggest that if administered as a combined regimen, it could potentially increase efficacy. , suggesting a pilot clinical trial to test this hypothesis. *(Arijs et al.Gut,67:43-52,2018;Gene Exp (Ression Omnibus database accession number GSE73661)

[0180] equivalent Those skilled in the art will be able to derive, using no more than routine experimentation, the specific embodiments of the invention described herein. It is likely that many equivalents of All of the foregoing are intended to be encompassed by the following claims. The contents of all references, patents, and published patent applications are incorporated herein by reference. do.

[0181] Table 2

[0182] Table 3

[0183] Table 4

[0184] Table 5

[0185] Table 6

[0186] Table 7

[0187] Table 8

Claims

1. 1. A method of treating a human patient in need thereof, said method comprising administering to said human patient α4 The method comprises administering a β7 inhibitor and an IL-23 inhibitor.

2. 2. The method of claim 1, wherein the α4β7 inhibitor is an anti-α4β7 antibody.

3. 3. The method of claim 2, wherein the anti-α4β7 antibody is humanized.

4. The anti-α4β7 antibody comprises a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3, a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO:2, and a DR2 domain; a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and ...2 domain set forth in SEQ ID NO: 8, a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and a CDR2 domain set forth in SEQ ID NO 6. The antibody of claim 2 or 3, comprising a light chain variable region comprising a CDR1 domain according to claim 6. method.

5. 1. A method of treating a human patient in need thereof, said method comprising administering to said human patient an anti-cancer drug. administering an α4β7 antibody and an IL-23 inhibitor; The anti-α4β7 antibody is an IgG1 antibody; and the CDR3 domain set forth in SEQ ID NO:

4. comprising a CDR2 domain set forth in SEQ ID NO:3 and a CDR1 domain set forth in SEQ ID NO:2 A heavy chain variable region comprising: a CDR3 domain set forth in SEQ ID NO: 8; a CDR set forth in SEQ ID NO: 7; a light chain variable region comprising a CDR1 domain set forth in SEQ ID NO: 6, and a CDR2 domain set forth in SEQ ID NO: 7; method.

6. The method of claim 1 or 5, wherein the human patient has an autoimmune disease.

7. 7. The method of claim 6, wherein the autoimmune disease is arthritis or psoriasis.

8. The autoimmune disease is selected from the group consisting of rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, and axial spondylitis. The method of claim 6, wherein the treatment is arthritis.

9. 10. The method of claim 1 or 5, wherein the human patient has inflammatory bowel disease (IBD).

10. 10. The method of claim 1 or 5, wherein the human patient has an autoimmune disease and an inflammatory bowel disease. method.

11. The method according to claim 9 or 10, wherein the IBD is ulcerative colitis or Crohn's disease. Law.

12. 12. The method of claim 11, wherein the ulcerative colitis is moderately to severely active ulcerative colitis. method.

13. 12. The method of claim 11, wherein the Crohn's disease is moderately to severely active Crohn's disease. 。

14. 14. The method according to claim 1, wherein the anti-α4β7 antibody is administered before the IL-23 inhibitor. The method according to any one of claims 1 to 4.

15. 14. The method according to claim 1, wherein the anti-α4β7 antibody is administered after the IL-23 inhibitor. The method according to any one of claims 1 to 4.

16. 13. The method of claim 1, wherein the anti-α4β7 antibody is administered concomitantly with the IL-23 inhibitor.

10. The method according to any one of the preceding claims.

17. The anti-α4β7 antibody comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:

1.

17. The method of claim 2, further comprising the step of:

10. The method according to any one of claims 1 to 9.

18. The method according to any one of claims 2 to 17, wherein the anti-α4β7 antibody is a humanized antibody. Law.

19. The method according to any one of claims 2 to 18, wherein the anti-α4β7 antibody is vedolizumab. method.

20. The human patient receives an initial dose of 300 mg of the anti-α4β7 antibody at week 0, followed by a 300 mg dose at week 2. A second administration of 300 mg of the anti-α4β7 antibody was administered at week 6, followed by a second administration of 300 mg of the anti-α4β7 antibody at week 6.

20. The method of any one of claims 2 to 19, wherein a third dose of 0.00 mg is administered.

21. Furthermore, starting 8 weeks after the third administration, the human patient is administered the anti-α4β7 antibody every 8 weeks.

21. The method of claim 20, comprising administering 300 mg of the compound.

22. Additionally, if the human patient does not show clinical improvement, the human patient is examined every four weeks.

22. The method of claim 21, comprising administering 300 mg of the anti-α4β7 antibody.

23. The human patient has ulcerative colitis or Crohn's disease, and the clinical improvement is clinical remission.

23. The method of claim 22, wherein the solution is

24. Furthermore, starting 8 weeks after the third administration, the human patient is administered the anti-α4β7 antibody every 4 weeks.

21. The method of claim 20, comprising administering 300 mg of the compound.

25. 25. The method of claim 20, wherein the anti-α4β7 antibody is administered intravenously. method.

26. Furthermore, the human patient receives two doses of the anti-α4β7 antibody every two weeks starting eight weeks after the third dose.

21. The method of claim 20, comprising administering 108 mg of the compound.

27. The human patient receives an initial dose of 300 mg of the anti-α4β7 antibody at week 0, followed by a 300 mg dose at week 2. A second administration of 300 mg of the anti-α4β7 antibody was administered at week 6, followed by a second administration of 100 mg of the anti-α4β7 antibody at week 6. A third dose of 0.08 mg is administered, followed by a dose of 108 mg every two weeks thereafter. Item 20. The method according to any one of Items 2 to 19.

28. 28. The method of claim 26 or 27, wherein the 108 mg dose is administered subcutaneously.

29. 29. The method of claim 28, wherein the 108 mg dose is self-administered.

30. The IL-23 inhibitor is an antibody that binds to the p19 subunit of IL-23.

30. The method according to any one of claims 1 to 29.

31. The IL-23 inhibitor is an antibody that binds to the p40 subunit of IL-23.

30. The method according to any one of claims 1 to 29.

32. The IL-23 inhibitor is risankizumab, ustekinumab, guselkumab, or thiazolinone. The method of any one of claims 1 to 29, wherein the compound is rudrakizumab.

33. The method according to any one of claims 1 to 27, wherein the IL-23 inhibitor is an antibody that binds to IL-23R. The method according to any one of claims 1 to 10.

34. The patient has a non- The method according to any one of claims 1 to 33, characterized as a responder or non-remitter. Law.

35. The patient has serum I at the start of treatment with an α4β7 inhibitor or 6 weeks after the start of treatment.

34. The method according to claim 1, characterized in that the level of L-22 is elevated. The method described.

36. The method of claim 34 or 35, wherein the α4β7 inhibitor is an anti-α4β7 antibody.

37. 37. The method of claim 36, wherein the anti-α4β7 antibody is vedolizumab.

38. The level of IL-22 in the serum of the human patient is determined by the initiation of treatment with the anti-α4β7 antibody. from the start of treatment with the anti-α4β7 antibody to 6 weeks after the start of treatment; or Six to ten weeks after the start of treatment with the anti-α4β7 antibody, there was no or only a 2-fold decrease The method of claim 5, wherein the ratio is reduced by less than one or by less than one-third.

39. the patient has a serum It has been characterized as having elevated levels of IL-22, with serum IL-22 levels , no decrease at all from the start to 10 weeks, from the start to 6 weeks, or from 6 weeks to 10 weeks, 6. The method of claim 5, wherein the reduction is less than two-fold or less than three-fold.

40. The patient has a decreased level of IL-22 and / or STAT5A compared to a control level.

10. The method of claim 1 or 5, characterized as having an increase.

41. The control level may be a subject not suffering from IBD, a healthy subject, a non-inflammatory subject from the patient.

41. The method of claim 40, at one or more levels of intestinal tissue or non-colonic tissue.

42. The patient's IL-22 and / or STAT5A levels are measured using the anti-α4β7 antibody.

42. The method of claim 40 or 41, wherein the measurement is performed before or on the first day of treatment.

43. The patient's IL-22 and / or STAT5A levels are measured using the anti-α4β7 antibody. The measurement is performed 1, 2, 3, 4, 5, 6, 7, 8, 9, and / or 10 days before the treatment.

43. The method of claim 42.

44. wherein said patient's IL-22 and / or STAT5A nucleic acid and / or protein levels The method according to any one of claims 35 to 42, wherein the .alpha.-to-.alpha.

45. the IL-22 and / or STAT5A levels are increased by 5%, 1%, or ０％、１５％、２０％、２５％、３０％、４０％、５０％、６０％、７０％、８０％、９ 41. The method of claim 40, wherein the increase is 0%, 100%, or more.

46. 1. A method of treating inflammatory bowel disease in a patient in need thereof, comprising administering an anti-α4 administering to said patient a β7 antibody and an antibody that binds to the p19 subunit of IL-23. wherein the anti-α4β7 antibody is administered to the human patient at week 0 by administering 300 ml of the anti-α4β7 antibody. g, followed by a second dose of 300 mg of the anti-α4β7 antibody at week 2, and a second dose of 300 mg of the anti-α4β7 antibody at week 6. a third administration of 300 mg of the anti-α4β7 antibody, followed by a second administration of 300 mg of the anti-α4β7 antibody 8 weeks after the third administration. receive a 300 mg dose of the anti-α4β7 antibody every week; The anti-α4β7 antibody comprises a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3, a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO:2, and a DR2 domain; a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and ...2 domain set forth in SEQ ID NO: 8, a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and a CDR2 domain set forth in SEQ ID NO a light chain variable region comprising a CDR1 domain set forth in no. 6; The patient has a decreased level of IL-22 and / or STAT5A compared to a control level. The method is characterized as having an increase.

47. 1. A method of treating inflammatory bowel disease in a patient in need thereof, comprising administering an anti-α4 administering to said patient a β7 antibody and an antibody that binds to the p40 subunit of IL-23. wherein the anti-α4β7 antibody is administered to the human patient at week 0 by administering 300 ml of the anti-α4β7 antibody. g, followed by a second dose of 300 mg of the anti-α4β7 antibody at week 2, and a second dose of 300 mg of the anti-α4β7 antibody at week 6. a third administration of 300 mg of the anti-α4β7 antibody, followed by a second administration of 300 mg of the anti-α4β7 antibody 8 weeks after the third administration. receive a 300 mg dose of the anti-α4β7 antibody every week; The anti-α4β7 antibody comprises a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3, a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO:2, and a DR2 domain; a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and ...2 domain set forth in SEQ ID NO: 8, a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and a CDR2 domain set forth in SEQ ID NO a light chain variable region comprising a CDR1 domain set forth in no. 6; The patient has a decreased level of IL-22 and / or STAT5A compared to a control level. The method is characterized as having an increase.

48. 1. A method of treating inflammatory bowel disease in a patient in need thereof, comprising administering an anti-α4 administering to said patient a β7 antibody and an antibody that binds to the p19 subunit of IL-23. wherein the anti-α4β7 antibody is administered by an initial administration of 300 mg of the anti-α4β7 antibody at week 0, followed by In the second week, a second dose of 300 mg of the anti-α4β7 antibody was administered, followed by a second dose of 300 mg of the anti-α4β7 antibody in the sixth week. A third dose of 108 mg of 4β7 antibody followed by 108 mg doses every 2 weeks thereafter. Given, The anti-α4β7 antibody comprises a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3, a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO:2, and a DR2 domain; a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and ...2 domain set forth in SEQ ID NO: 8, a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and a CDR2 domain set forth in SEQ ID NO a light chain variable region comprising a CDR1 domain set forth in no. 6; The patient has a decreased level of IL-22 and / or STAT5A compared to a control level. The method is characterized as having an increase.

49. 1. A method of treating inflammatory bowel disease in a patient in need thereof, comprising administering an anti-α4 administering to said patient a β7 antibody and an antibody that binds to the p40 subunit of IL-23. wherein the anti-α4β7 antibody is administered by an initial administration of 300 mg of the anti-α4β7 antibody at week 0, followed by In the second week, a second dose of 300 mg of the anti-α4β7 antibody was administered, followed by a second dose of 300 mg of the anti-α4β7 antibody in the sixth week. A third dose of 108 mg of 4β7 antibody followed by 108 mg doses every 2 weeks thereafter. Given, The anti-α4β7 antibody comprises a CDR3 domain set forth in SEQ ID NO: 4, a CDR4 domain set forth in SEQ ID NO: 3, a heavy chain variable region comprising a CDR1 domain set forth in SEQ ID NO:2, and a DR2 domain; a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and ...2 domain set forth in SEQ ID NO: 8, a CDR3 domain set forth in SEQ ID NO: 8, a CDR2 domain set forth in SEQ ID NO: 7, and a CDR2 domain set forth in SEQ ID NO a light chain variable region comprising a CDR1 domain set forth in no. 6; The patient has a decreased level of IL-22 and / or STAT5A compared to a control level. The method is characterized as having an increase.

50. The control level may be a subject not suffering from IBD, a healthy subject, a non-inflammatory subject from the patient.

50. The method of claim 46, wherein the method is performed at one or more levels of intestinal tissue or non-colonic tissue.

1. The method according to claim 1.

Images