Safe and effective treatment of ulcerative colitis with anti-IL12 / IL23 antibodies

Abstract

The present invention describes methods and compositions for the clinically proven, safe and effective treatment of ulcerative colitis, particularly moderately to severely active ulcerative colitis, in patients who have an inadequate response to or poor tolerance to conventional or existing therapies by intravenous and / or subcutaneous administration of anti-IL-12 / IL-23p40 antibodies.

Description

[Technical Field]

[0001] (Reference to electronically submitted sequence listings) This application includes a sequence listing filed electronically via EFS-Web on January 26, 2021, in ASCII format, with the filename "JBI6165WOPCT2Sequence Listing.txt", and having a size of 15 kilobytes. The sequence listing filed via EFS-Web is part of this specification and is incorporated herein by reference in its entirety.

[0002] (Field of Invention) The present invention relates to a method for providing a clinically proven safe and clinically proven effective treatment for ulcerative colitis, particularly moderate to severe active ulcerative colitis, in patients who have an insufficient response to or poor tolerance to conventional or existing therapies involving intravenous and / or subcutaneous administration of anti-IL-12 / IL-23p40 antibodies. [Background technology]

[0003] Inflammatory bowel disease (IBD), including ulcerative colitis (UC), is a chronic, relapsing disease characterized by destructive inflammation and epithelial damage to the gastrointestinal (GI) tract (Baumgart and Sandborn, J Clin Invest. 98:1010-1020 (1996); Danese and Fiocchi, N Engl J Med. 365:1715-1725 (2011)). The prevalence of UC in the United States is estimated to be 205-240 cases per 100,000 people, or 9-12 cases per 100,000 people (Tally et al., Am J Gastroenterol. 106 Suppl 1:S2-S25 (2011)). The estimated prevalence of ulcerative colitis (UC) in Europe is approximately 1,000,000 people (Loftus, Gastroenterology. 126(6):1504-1517 (2004); Looftus, Gastoenterol Clin N Am. 31:1-20 (2002)). The etiology of UC is unknown. However, it is thought that an abnormal immune response to the contents of the intestines, including intestinal bacteria, causes the disease in genetically predisposed individuals (Geremia et al., Autoimmun Rev. 13:3-10 (2014)). Dysregulated congenital and adaptive immune pathways contribute to abnormal enteritis in IBD, and cytokines including interleukin (IL)-12, interferon-gamma (IFNγ), and IL-23 are involved in the pathogenesis of ulcerative colitis (Geremia et al., Autoimmune Rev. 2014; 13: 3-10; Neurath, Nat Rev Immunol. 14(5): 329-42 (2014)).

[0004] The involvement of the IL-12 / IL-23 pathway in the pathogenesis of IBD is well established, and the important role of the IL-12 / IL-23 pathway in enteritis has been elucidated in colitis (Ahern et al., Immunity. 33(2):279-288 (2010); Investigator's Brochure: STELARA(registered trademark) (ustekinumab), edition 18. Janssen Research & Development, LLC (2017), Uhlig et al., Immunity. 25:309 318 (2006); Yen et al., J Clin Invest. 116(5):1310-1316 (2006)). Early studies have shown that treatment with anti-IFNγ (Berg et al., J Clin Invest. 98:1010-1020 (1996); Davidson et al., J Immunol. 161:3143-3149 (1998)) or anti-IL-12p40 monoclonal antibody (mAb) prevents disease in experimental models of colitis, suggesting an important role for type 1 helper T (Th-1) cells that promote intestinal inflammation (Neurath et al., J Exp Med. 182(5):1281-1290 (1995)). Genome-wide association studies have identified several human loci in the IL-12 / 23 pathway associated with increased susceptibility to UC, including IL-23 R and IL-12 B (Anderson et al., Nat Genet. 43(3):246-252 (2011); Brant et al., Clin Gastroenterol Hepatol. 11(1):22-26 (2013)). Subjects with active UC have been shown to have significantly more IL-23, IL-22, IL-22R1, and p-STAT3 positive cells than subjects with inactive UC and normal controls (Yu et al., World J Gastroenterol. 19(17):2638-2649 (2013)).

[0005] The currently approved biological therapies for the treatment of UC are either tumor necrosis factor (TNF) or integrin inhibitors (Colombel et al., Gastroenterology. 132:52-65 (2007); Hanauer et al., Lancet. 359:1541-1549 (2002); Sandborn et al., N Engl J Med. 369:711-721 (2013); Sandborn et al., Gastroenterology. 142:257-265 (2012)). However, of all currently approved therapies, only vedolizumab has demonstrated efficacy in patients with an inadequate response to anti-TNF (i.e., primary or secondary non-response) or poor tolerance to anti-TNF (Feagan et al., N Engl J Med. 369:699710 (2013)). Anti-TNF therapy carries safety risks associated with immunosuppression, and not all patients respond appropriately to such treatments. Furthermore, inadequate responses and intolerances, as observed with anti-TNF therapy, have been identified in patients receiving vedolizumab for the treatment of UC. Therefore, there remains an unmet need for novel therapies with alternative mechanisms of action.

[0006] Upon investigation, it was found that biological therapies currently approved for the treatment of UC also show efficacy in Crohn's disease (Sandborn et al., Gastroenterology. 135(4):1130-1141 (2008)). Numerous pieces of evidence suggest that inflammatory bowel disease (UC and Crohn's disease) is mediated by Th1 or Th17 cells with a strong contribution from inflammatory cytokines, IL-12 and IL-23. Ustekinumab (STELARA®) is a fully human immunoglobulin G1 mAb against human IL-12 / 23p40, which inhibits the bioactivity of IL-12 and IL-23 by preventing them from interacting with their cell surface IL-12Rβ1 receptor protein (Investigator's Brochure: STELARA® (ustekinumab), edition 18. Janssen Research & Development, LLC (2017)). This mechanism of action allows ustekinumab to effectively neutralize IL-12(Th1)- and IL-23(Th17)-mediated cellular responses. Ustekinumab is approved for marketing worldwide, including in North America, Europe, South America, and the Asia Pacific region, for the treatment of adults with moderate to severe active Crohn's disease (initial approval for Crohn's disease was received on November 11, 2016), adults with moderate to severe plaque psoriasis, or adults with active psoriatic arthritis, and children (12-17 years) with moderate to severe plaque psoriasis.

[0007] The efficacy and safety of intravenous (IV) ustekinumab as induction therapy in Crohn's disease have been evaluated in clinical trials CRD3001 and CRD3002. Trial CRD3001 evaluated subjects with prior failure or intolerance to one or more TNF antagonists, while CRD3002 evaluated subjects with a history of inadequate response or intolerance to corticosteroids or immunomodulators, but without a history of inadequate response or intolerance to TNF antagonists. These trials evaluated two IV doses: a fixed IV dose of 130 mg (approximately 2 mg / kg) was selected as the low-dose group, and a dose based on body weight range of approximately 6 mg / kg IV (body weight ≤ 55 kg: ustekinumab 260 mg; body weight > 55 and ≤ 85 kg: ustekinumab 390 mg; body weight > 85 kg: ustekinumab 520 mg) was selected as the high-dose group. In both trials, ustekinumab demonstrated clinically significant efficacy compared to placebo and was well-tolerated with a favorable safety profile. [Overview of the project] [Problems that the invention aims to solve]

[0008] Prior to the present invention, no studies using ustekinumab for UC had been conducted, and there is a need in the art for an improved method of treating UC, particularly moderate to severe active UC, in subjects who have previously been unsuccessful with or poorly tolerated by both biological therapies and other conventional therapies, or who have shown corticosteroid dependence. [Means for solving the problem]

[0009] This application relates to clinically proven safe methods and compositions, as well as clinically proven effective methods and compositions, for the treatment of moderate to severe active ulcerative colitis (UC) in subjects who have an inadequate response to or poor tolerance to conventional or existing therapies, by administration of an anti-IL-12 / IL-23p40 antibody, thereby addressing a clearly unmet medical need in this subject population.

[0010] In one general embodiment, the present application relates to a clinically proven safe and clinically proven effective method for treating moderate to severe active ulcerative colitis (UC) in subjects in need thereof, the method comprising administering to a subject a pharmaceutical composition comprising a safe and effective amount of anti-IL-12 / IL-23p40 antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the complementarity-determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, the CDRH2 amino acid sequence of SEQ ID NO: 2, and the CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprising the complementarity-determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO: 4, the CDRL2 amino acid sequence of SEQ ID NO: 5, and the CDRL3 amino acid sequence of SEQ ID NO: 6.

[0011] In a particular embodiment, the anti-IL-12 and / or anti-IL-23 antibody is administered intravenously to the subject, preferably at week 0, at a dose of approximately 6.0 mg / kg of the subject's body weight or 130 mg per administration.

[0012] In a particular embodiment, anti-IL-12 and / or anti-IL-23 antibodies are administered intravenously or subcutaneously to the subject, preferably at 8 weeks, at a dose of approximately 6.0 mg / kg of the subject's body weight or 90 mg per administration.

[0013] Preferably, subjects treated by the methods according to embodiments of this application had an inadequate response to or poor tolerance to conventional or existing therapies. In some embodiments, subjects had previously failed to or poorly tolerated biological therapies such as anti-TNF and / or vedolizumab. In some embodiments, subjects had previously failed to or poorly tolerated non-biological therapies such as treatment with corticosteroids, azathioprine (AZA), and / or 6-mercaptopurine (6MP). In some embodiments, subjects exhibited corticosteroid dependence.

[0014] In another general aspect, the present application relates to a clinically proven safe and clinically proven effective method for treating moderate to severe active ulcerative colitis (UC) in patients in need, wherein the method is The target population will receive an intravenous administration of a pharmaceutical composition containing an anti-IL-12 / IL-23p40 antibody at a dose of approximately 6.0 mg / kg of the target population or 130 mg of antibody per administration during week 0 of treatment. The treatment involves subcutaneously administering a pharmaceutical composition containing an anti-IL-12 / IL-23p40 antibody at a dose of 90 mg of antibody per administration during the 8th week of treatment. The antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the complementarity-determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, the CDRH2 amino acid sequence of SEQ ID NO: 2, and the CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprising the complementarity-determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO: 4, the CDRL2 amino acid sequence of SEQ ID NO: 5, and the CDRL3 amino acid sequence of SEQ ID NO: 6. The subjects had previously failed or had poor tolerance to at least one therapy selected from the group consisting of anti-TNF, vedolizumab, corticosteroids, azathioprine (AZA), and 6-mercaptopurine (6MP), or the subjects showed corticosteroid dependence.

[0015] In certain embodiments, the method of the present application comprises administering intravenously (IV) and / or subcutaneously (SC) to a subject a pharmaceutical composition comprising an anti-IL-12 and / or anti-IL-23 antibody or antigen-binding fragment comprising (i) the heavy chain variable region amino acid sequence of SEQ ID NO: 7 and (ii) the light chain variable region amino acid sequence of SEQ ID NO: 8.

[0016] In a particular embodiment, the method of the present application comprises administering to a subject intravenously (IV) and / or subcutaneously (SC) a pharmaceutical composition comprising ustekinumab, an anti-IL-12 / 23p40 antibody, comprising (i) the heavy chain amino acid sequence of SEQ ID NO: 10 and (ii) the light chain amino acid sequence of SEQ ID NO: 11.

[0017] In a particular embodiment, the IV dose for week 0 is approximately 6.0 mg / kg. For example, the IV dose is 260 mg for subjects weighing 35 kg or more but 55 kg or less, 390 mg for subjects weighing more than 55 kg but 85 kg or less, and 520 mg for subjects weighing more than 85 kg.

[0018] In certain embodiments, subjects are responders to treatment by a method according to one embodiment of the present application, and are identified as having at least one of the following, measured preferably for 92 weeks after initial treatment and maintenance administration: (1) clinical remission based on at least one of the World Application and the US Application; (2) endoscopic healing; (3) clinical response; (4) change from baseline in the Inflammatory Bowel Disease Questionnaire (IBDQ) score; (5) mucosal healing; (6) decrease from baseline in the Mayo score; and (7) normalization of one or more biomarkers selected from the group consisting of C-reactive protein, fecal lactoferrin, and fecal calprotectin. Preferably, at least one of (1) to (7) above is identified from subjects by week 16 of treatment, more preferably by week 8 or week 4, and most preferably by week 2.

[0019] In certain embodiments, the present invention provides a clinically proven safe and clinically proven effective method for treating moderate to severe active UC in subjects, who are identified as responders to antibody therapy and who have a statistically significant improvement in disease activity, as determined by endoscopic cure with a Mayo endoscopic subscore of 0 or 1, by the 8th week of antibody therapy.

[0020] In other embodiments, the present invention provides a clinically proven safe method and a clinically proven effective method for treating moderately to severely active UC in a subject, where the subject is a responder to antibody treatment and is identified as having a statistically significant improvement in disease activity determined by the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) score of ≤ 4 by week 8 of antibody treatment.

[0021] In certain embodiments, the subject exhibits a clinical response determined by a decrease of 30% or more and ≥ 3 points from baseline in the Mayo score and a decrease from baseline in the rectal bleeding subscore of ≥ 1 point by week 8 of antibody treatment, or a rectal bleeding subscore of 0 or 1.

[0022] In other embodiments, the maintenance dose of the anti-IL-12 / IL-23p40 antibody is administered every 8 weeks or every 12 weeks after treatment at week 8, and the clinical response is maintained by the subject for at least 44 weeks.

[0023] In certain embodiments, the present invention provides a clinically proven safe method and a clinically proven effective method for treating moderately to severely active UC in a subject, and a second treatment is preferably administered to a subject identified as a non-responder to initial treatment by a different route of administration than the initial treatment. For example, a subject identified as a non-responder to initial treatment by IV administration of an antibody or antibody fragment can be treated with subsequent subcutaneous administration of an antibody or antibody fragment according to an embodiment of the present invention.

[0024] In certain embodiments, the present application provides a method for treating moderate to severe active UC in a subject, and the anti-IL-12 and / or anti-IL-23 antibody for use in intravenous administration is in a pharmaceutical composition comprising a solution containing 10 mM L-histidine, 8.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80, 0.4 mg / mL L-methionine, and 20 μg / mL disodium EDTA dihydrate at pH 6.0.

[0025] In certain embodiments, the present application provides a clinically proven safe and clinically proven effective method for treating moderate to severe active UC in a subject, and the anti-IL-12 and / or anti-IL-23 antibody for use in subcutaneous administration is in a pharmaceutical composition comprising a solution containing 6.7 mM L-histidine, 7.6% (w / v) sucrose, 0.004% (w / v) polysorbate 80 at pH 6.0.

[0026] In certain embodiments, the present application further provides a method that further comprises administering to the subject one or more additional drugs used to treat UC. In a preferred embodiment, the additional drug is selected from the group consisting of an oral 5-aminosalicylate (5-ASA) compound, an oral corticosteroid, an immunomodulator, 6-mercaptopurine (6-MP), azathioprine (AZA), or methotrexate (MTX).

[0027] Another aspect of the present application includes a pharmaceutical composition comprising an anti-IL-12 and / or anti-IL-23 antibody for use in a clinically proven safe and clinically proven effective method for treating moderate to severe active UC in a subject, as well as a method for preparing the composition and a kit comprising the pharmaceutical composition.

[0028] In certain embodiments, a kit useful in the method of the present invention comprises at least one of the pharmaceutical composition for intravenous administration of the present invention and the pharmaceutical composition for subcutaneous administration of the present invention. In other embodiments, the kit comprises both the pharmaceutical composition for intravenous administration and the pharmaceutical composition for subcutaneous administration of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The above-mentioned "Summary of the Invention" and the following "Modes for Carrying Out the Invention" will be better understood when read in conjunction with the attached drawings. It should be understood that the present invention is not limited to the exact embodiments shown in the drawings. [Figure 1] This is a diagram illustrating the test design for the introductory and maintenance trials of the Phase 3 program design. Abbreviations: W8 = Week 8, W16 = Week 16, LTE = Long-term continuous administration. [Figure 2] This figure shows the maintenance test for the Phase 3 program design. [Figure 3] This figure shows the distribution of subjects in the CNTO1275UC03001 group based on administration during the maintenance phase from week 0 to week 96. [Figure 4] This figure shows the distribution of subjects in the non-randomized CNTO1275USO3001 group based on administration during the maintenance phase from week 0 to week 96. [Figure 5] This figure shows the proportion of patients who achieved symptomatic remission up to week 92 or until dose adjustment in maintenance studies administered with LTE (CNTO1275USO3001). [Figure 6] This figure shows the average daily dose of corticosteroids (in prednisone equivalent) from week 0 to week 92 during the maintenance phase baseline for subjects who received corticosteroids other than budesonide and beclomethasone dipropionate. [Figure 7] This figure shows the number of subjects who achieved symptomatic remission over the period up to week 92. Here, dose adjustments are made because randomized subjects in the maintenance phase who received long-term continued administration are not considered to have failed the treatment. [Figure 8] This figure shows the number of subjects who achieved symptomatic remission over the period up to week 92. Here, all subjects were randomized at week 0 of the maintenance phase and were not considered ineffective due to dose adjustment. [Modes for carrying out the invention]

[0030] In the background art and throughout this specification, various publications, articles, and patents are cited or referenced, and each of these references is incorporated herein by reference in its entirety. The considerations of documents, operations, materials, devices, articles, etc., included herein are for the purpose of providing context for the present invention. Such considerations do not constitute an endorsement that any or all of these things constitute part of the prior art to any invention disclosed or claimed.

[0031] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art in which the present invention pertains. Otherwise, any particular term used herein has the meaning set forth herein. All patents, published patent applications and publications referenced herein are incorporated by reference as if they were included herein in their entirety.

[0032] When used herein and in the appended claims, it should be noted that the singular forms "a," "an," and "the" refer to multiple objects unless otherwise clearly indicated by the context.

[0033] Unless otherwise stated, the term “at least” preceding a set of elements should be understood to refer to all of those elements. Those skilled in the art will recognize or confirm many equivalents to the specific embodiments of the invention described herein by simply using ordinary experimental procedures. Such equivalents are intended to be encompassed by the invention.

[0034] Throughout this specification and the following claims, unless contextually required, the term “comprise” and variations such as “comprises” and “comprising” will be understood to mean that they include a specified integer or step or group of integers or steps, but not any other integer or step or group of integers or steps. As used herein, the term “comprising” may be replaced with the terms “containing” or “including,” or, as sometimes used herein, with the term “having.”

[0035] As used herein, “consisting of” excludes any element, step, or component not specified in the elements of the claims. As used herein, “consisting essentially of” does not exclude materials or steps that do not substantially affect the basic and novel features of the claims. As used herein in relation to aspects or embodiments of the present invention, any of the above terms “comprising,” “containing,” “including,” and “having” may be replaced with the terms “consisting of” or “consisting essentially of” to change the scope of this disclosure.

[0036] When used herein, the connecting term "and / or" between multiple enumerated elements is understood to encompass both individual and combined options. For example, when two elements are connected by "and / or," the first option refers to the applicability of the first element without the second element. The second option refers to the applicability of the second element without the first element. The third option refers to the applicability of the first and second elements together. Any one of these options is included in the meaning and therefore satisfies the requirements of the term "and / or" when used herein. The simultaneous applicability of two or more of the options is also included in the meaning and therefore satisfies the requirements of the term "and / or."

[0037] As used herein, “Subject” means any animal, preferably a mammal, most preferably a human, that is treated or has been treated by the methods according to embodiments of the present invention. As used herein, the term “mammal” encompasses all mammals. Examples of mammals, but not limited to, include, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys or apes, non-human primates (NHPs), humans, and more preferably humans.

[0038] As used herein, the term “combined use” refers to the use of multiple therapeutic agents in relation to the administration of two or more therapeutic agents to a subject. The use of the term “combined use” is not limited to the order in which the therapeutic agents are administered to the subject. For example, the first therapeutic agent (e.g., a composition described herein) may be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 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 prior), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 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 prior) the administration of the second therapeutic agent to the subject.

[0039] As used herein, “anti-IL-12 antibody,” “anti-IL-23 antibody,” “anti-IL-12 / 23p40 antibody,” or “IL-12 / 23p40 antibody” refers to a monoclonal antibody (mAb) or its antigen-binding fragment that binds to the 40 kDa (p40) subunit shared by the cytokines interleukin-12 and interleukin-23 (IL-12 / 23p40). This antibody can affect at least one of the following IL-12 / 23 activities or functions, including but not limited to RNA, DNA, or protein synthesis, IL-12 / 23 release, IL-12 / 23 receptor signaling, membrane IL-12 / 23 cleavage, IL-12 / 23 activity, IL-12 / 23 production, and / or synthesis.

[0040] The term “antibody” is further intended to encompass antibodies, their digestive fragments, identified parts, and variants, including antibody mimetic drugs, or parts of antibodies or identified fragments or portions thereof that mimic the structure and / or function of antibodies, such as single-chain antibodies and their fragments. Functional fragments include antigen-binding fragments that bind to mammalian IL-12 / 23. Examples include, but are not limited to, Fab (e.g., by papain digestion), Fab' (e.g., by pepsin digestion and partial reduction), and F(ab')2 (e.g., by pepsin digestion), facb (e.g., by plasmin digestion), pFc' (e.g., by pepsin or plasmin digestion), Fd (e.g., by pepsin digestion, partial reduction and reassembly), Fv or scFv (e.g., by molecular biological techniques) fragments, antibody fragments capable of binding to IL-12 / 23 or a portion thereof are included in the present invention (see, for example, Colligan, Immunology above).

[0041] Such fragments can be produced by enzymatic cleavage, synthesis, or recombination techniques known in the art and / or described herein. Antibodies can also be produced in various cleavage types using antibody genes in which one or more stop codons are introduced upstream of the native stop site. For example, a combination of genes encoding the F(ab') double helix region is the C of the heavy chain. H The antibody can be designed to include a DNA sequence encoding one domain and / or a hinge region. Various parts of the antibody can be chemically linked using conventional techniques, or it can be prepared as a seamless, continuous protein using genetic engineering techniques.

[0042] As used herein, the term “human antibody” means substantially all parts of a protein (e.g., CDR, framework, C). L , C H Domain (for example, C H 1, C H 2, C H 3) Hinge (V L , V HHuman antibodies refer to antibodies that are substantially non-immunogenic in humans, possessing only minor sequence changes or mutations. "Human antibodies" can also be antibodies derived from or closely matching human germline immunoglobulin sequences. Human antibodies may contain amino acid residues not encoded by germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutations in vitro, or by somatic mutations in vivo). In many cases, this means that human antibodies are substantially non-immunogenic in humans. Human antibodies are grouped based on the similarity of their amino acid sequences. Therefore, sequence similarity searches can be used to select antibodies with similar linear sequences as templates for creating human antibodies. Similarly, antibodies whose names include primates (monkeys, baboons, chimpanzees, etc.), rodents (mice, rats, rabbits, guinea pigs, hamsters, etc.), and other mammals specify antibodies specific to such species, subgenus, genus, subfamily, and family. Furthermore, chimeric antibodies may include any combination of the above. Such changes or mutations may, and preferably, preserve or reduce immunogenicity in humans or other species compared to unmodified antibodies. Therefore, human antibodies are different from chimeric antibodies or humanized antibodies.

[0043] It has been noted that human antibodies can be produced by non-human animals, or by prokaryotic or eukaryotic cells, that can express functionally reconstituted human immunoglobulin (e.g., heavy chain and / or light chain) genes. Furthermore, if a human antibody is a single-chain antibody, it may contain linkapeptides not found in naturally occurring human antibodies. For example, Fv may contain linkapeptides such as 2 to about 8 glycine or other amino acid residues that connect the variable regions of the heavy chain and the variable regions of the light chain. Such linkapeptides are considered to be of human origin.

[0044] Anti-IL-12 / 23p40 antibodies (also referred to as IL-12 / 23p40 antibodies) (or antibodies against IL-23) useful in the methods and compositions of the present invention may optionally be characterized by high affinity binding to IL-12 / 23p40 and optionally, and preferably, low toxicity. Specifically, antibodies, identified fragments thereof, or variants of the present invention are useful in the present invention, in which individual components such as variable regions, constant regions, and frameworks individually and / or collectively optionally, and preferably, have low immunogenicity. Antibodies that can be used in the present invention are preferably characterized by the ability to treat a subject for a long period of time with measurable symptom relief and low and / or tolerable toxicity. Low or tolerable immunogenicity, and / or high affinity, and other desirable properties may contribute to the therapeutic outcome obtained. "Low immunogenicity" is defined herein as a significant increase in the HAHA, HACA, or HAMA response in less than approximately 75%, preferably less than approximately 50%, of the subjects being treated, and / or an increase in low titers (less than approximately 300, preferably less than approximately 100, as measured by a dual antigen enzyme immunoassay) in the subjects being treated (Elliott et al., Lancet 344:1125-1127 (1994), the whole of which is incorporated herein by reference). "Low immunogenicity" can also be defined as the expression rate of titration-level antibodies against anti-IL-12 antibodies in subjects being treated with anti-IL-12 antibodies, expressed in less than 25%, preferably less than 10%, of the subjects being treated at the recommended dose throughout the recommended course of treatment during the treatment period.

[0045] The terms “clinically proven efficacy” and “clinically proven effective” as used herein in the context of dose, administration regimen, treatment or method refer to the efficacy of a particular dose, administration, or administration regimen. Efficacy may be measured based on changes in the course of the disease in response to the agent of the present invention. For example, the anti-IL12 / 23p40 (e.g., ustekinumab) of the present invention is administered to a subject in an amount and time sufficient to produce improvement, preferably sustained improvement, in at least one indicator reflecting the severity of the disorder being treated. Various indicators reflecting the extent of the disease, illness or condition in the subject may be evaluated to determine whether the amount and time of treatment are sufficient. Such indicators include, for example, clinically recognized indicators of disease severity, symptoms, or the manifestation of the disorder in question. The degree of improvement is determined overall by a physician, who may make this determination based on signs, symptoms, biopsy, or other test results, or by employing questionnaires administered to the subject, such as quality of life questionnaires developed in relation to a given disease. For example, the anti-IL12 / 23p40 or anti-IL23 antibody of the present invention may be administered to achieve improvement in the condition of a subject associated with ulcerative colitis.

[0046] This improvement can be indicated by an improvement in the disease activity index, remission of clinical symptoms, or any other measure of disease activity. The previous disease index is the ulcerative colitis Mayo score. The Mayo score is an established and validated disease activity index for mild, moderate, and severe ulcerative colitis (UC), ranging from 0 to 12, calculated as the sum of four subscores: bowel movement frequency, rectal bleeding, endoscopic findings, and physician's overall assessment (PGA). A score of 3–5 indicates mild disease activity, a score of 6–10 indicates moderate disease activity, and a score of 11–12 indicates severe disease. The partial Mayo score, which is the Mayo score without the endoscopic subscore, is calculated as the sum of the subscores for bowel movement frequency, rectal bleeding, and physician's overall assessment, ranging from 0 to 9. The modified Mayo score, which is the Mayo score without PGA subscores, is calculated as the sum of bowel movement frequency, rectal bleeding, and endoscopic subscores, and ranges from 0 to 9. Other indicators of disease activity for UC include, for example, the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) score and the Bristol Stool Form Scale (BSFS) score. The UCEIS score provides a comprehensive assessment of the endoscopic severity of UC based on mucovascular patterns, bleeding, and ulceration (Travis et al., Gut. 61:535-542 (2012)). The score ranges from 3 to 11, with higher scores indicating more severe disease on endoscopy. The BSFS score is used to classify the morphology (or viscosity) of human feces into seven categories (Lewis and Heaton, Scand J Gastroenterol. 32(9):920-924 (1997)).

[0047] As used herein, the term “clinical response” refers, when relating to a subject’s response to drug administration, to a decrease from baseline in either a rectal bleeding subscore of ≥1 or a Mayo score of ≥30% and ≥3 points, accompanied by either a rectal bleeding subscore of 0 or 1.

[0048] The term "clinically proven safety," in the context of the dose, administration regimen, treatment, or method of the anti-IL-12 / IL-23p40 antibody (e.g., ustekinumab) of the present invention, refers to a favorable risk-benefit ratio with an acceptable frequency and / or acceptable severity of adverse events (referred to as adverse events, AEs, or treatment-emergent adverse events, TEAEs) occurring under treatment, compared to standard treatment or another comparator. As used herein, "adverse event," "adverse event occurring during treatment," and "adverse reaction" mean any harmful, undesirable, unintended, or unwanted sign or result related to or caused by the administration of a pharmaceutical composition or therapeutic agent. This is an undesirable medical event in a subject to which the drug has been administered. However, abnormal values ​​or observations are not reported as adverse events unless they are considered clinically significant by the principal investigator. As used herein, when referring to an adverse event, "clinically evident" means clinically significant as determined by a physician or principal investigator using criteria acceptable to those skilled in the art. If the adverse or undesirable consequences of an adverse event reach such a severity, regulatory authorities may consider the pharmaceutical composition or therapeutic agent unacceptable for the proposed use. Specifically, “safety” in relation to the dosage, administration regimen, or treatment with the anti-IL12 / 23p40 or anti-IL23 antibody of the present invention refers to the acceptable frequency and / or acceptable severity of adverse events associated with the administration of the antibody, where the cause is likely, highly likely, or very likely to be due to the use of the anti-IL12 / 23p40 or anti-IL23 antibody.

[0049] As used herein, unless otherwise specified, the term “clinically proven” (used independently or to modify the terms “safety” and / or “effectiveness”) means proven by clinical trials that meet the approval standards of the U.S. Food and Drug Administration, EMEA, or the corresponding national regulatory agency. For example, a clinical trial may be a randomized, double-blind trial of appropriate size used to clinically prove the efficacy of a drug.

[0050] When used herein, the dose of anti-IL-12 / IL-23p40 antibody in "mg / kg" units refers to the amount of milligrams of anti-IL-12 / IL-23p40 antibody per kilogram of body weight of the recipient.

[0051] Antibody production and generation according to the present invention At least one anti-IL-12 / 23p40 (or anti-IL-23) used in the method of the present invention can be optionally produced by cell lines, mixed cell lines, immortalized cells, or clonal populations of immortalized cells that are well known in the art. For example, see Ausubel, et al., ed., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, NY (1987-2001), Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, NY (1989), Harlow and Lane, "antibodies," a Laboratory Manual, Cold Spring Harbor, NY (1989), Colligan et al., eds., Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001), and Colligan et al., Current Protocols in Protein Science, John Wiley & Sons, NY, NY (1997-2001), each of which is incorporated herein by reference in its entirety.

[0052] Human antibodies specific to human IL-12 / 23p40 or IL-23 proteins or fragments thereof may be generated against suitable immunogenic antigens such as isolated IL-12 / 23p40 proteins, IL-23 proteins, and / or portions thereof (including synthetic molecules such as synthetic peptides). Antibodies of other specific or common mammals may similarly be generated. The preparation of immunogenic antigens and the production of monoclonal antibodies can be carried out using any suitable technique in consideration of this disclosure.

[0053] One approach involves using appropriate immortalized cell lines (e.g., Sp2 / 0, Sp2 / 0-AG14, NSO, NS1, NS2, AE-1, L.5, L243, P3X63Ag8.653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 SA5, U937, MLA 144, ACT IV, MOLT4, DA-1, JURKAT, WEHI, K-562, COS, RAJI, NIH 3T3, HL-60, MLA 144, NAMALWA, NEURO) Myeloma cell lines such as 2A, or heteromylomas, their fusion products, or any cells or fusion cells derived therefrom, or any other suitable cell lines known in the art) (see, for example, www.atcc.org, www.lifetech.com, etc.), but not limited to, isolated or cloned antibody-producing cells such as spleen, peripheral blood, lymph, tonsil, or other immune or B cell-containing cells, or as endogenous or heteronucleotides, recombinant or endogenous, viruses, bacteria, algae, Hybridomas are produced by fusing with any other cell expressing a constant or variable heavy or light chain, or a framework or CDR sequence, in any form such as prokaryotes, amphibians, insects, reptiles, fish, mammals, rodents, horses, sheep, goats, primates, eukaryotes, genomic DNA, cDNA, rDNA, mitochondrial DNA or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single, double or triple stranded, hybridized, or any combination thereof. See, for example, Chapter 2 of Ausubel and Colligan, Immunology. Both references are incorporated herein by reference in their entirety.

[0054] Antibody-producing cells can also be obtained from the peripheral blood of a human or other suitable animal immunized with the antigen of interest, preferably from the spleen or lymph nodes. Any other suitable host cells can also be used to express heterologous or endogenous nucleic acids encoding the antibodies of the present invention, the identified fragments, or their variants. Fusion cells (hybridoms) or recombinant cells can be isolated using selective culture conditions or other suitable known methods and cloned by limiting dilution, cell sorting, or other known methods. Cells producing antibodies with desired specificity can be selected by a suitable assay (e.g., ELISA).

[0055] Methods for selecting recombinant antibodies from peptide or protein libraries are included, but are not limited to, other suitable methods for producing or isolating antibodies with the required specificity (e.g., display libraries such as bacteriophages, ribosomes, oligonucleotides, RNA, cDNA, etc., e.g., Cambridge Antibody Technologies, Cambridgeshire, UK; MorphoSys, Martinsreid / Planegg, DE; Biovation, Aberdeen, Scotland, UK; BioInvent, Lund, Sweden; Dyax Corp., Enzon, Affymax / Biosite, Xoma, Berkeley, CA; Ixsys).(Available from) For example, European Patent No. 368,684, International Application PCT / GB91 / 01134, International Application PCT / GB92 / 01755, International Application PCT / GB92 / 002240, International Application PCT / GB92 / 00883, International Application PCT / GB93 / 00605, US Patent Application No. 08 / 350260 (5 / 12 / 94), International Application PCT / GB94 / 01422, International Application PCT / GB94 / 02662, International Application PCT / GB97 / 01835, (CAT / MRC), International Publication No. 90 / 14443, International Publication No. 90 / 14424, International Publication No. 90 / 14430, International Application PCT / US94 / 1234, International Publication No. 92 / 18619, International Publication No. 96 / 07754 (Scripps), International Publication No. 96 / 13583, International Publication No. 97 / 08320 (MorphoSys), International Publication 95 / 16027 (BioInvent), International Publication 88 / 06630, International Publication 90 / 3809 (Dyax), U.S. Patent No. 4,704,692 (Enzon), International Application PCT / US91 / 02989 (Affymax), International Publication 89 / 06283, European Patent No. 371998, European Patent No. 550 Patent No. 400, (Xoma), European Patent No. 229046, International Application PCT / US91 / 07149 (Ixsys), or Probabilistically Produced Peptides or Proteins - U.S. Patents No. 5723323, 5763192, 5814476, 5817483, 5824514, 5976862, International Publication No. 86 / 05803, European Patent No. 590 689 (Ixsys, a predecessor to Applied Molecular Evolution (AME), each of which is incorporated herein by reference in whole) or relies on immunization of transgenic animals capable of producing a repertoire of human antibodies known in the art and / or described herein (e.g., SCID mice, Nguyen et al., Microbiol.Immunol. 41:901-907 (1997); Sandhu et al., Crit.Rev.Biotechnol. 16:95-118 (1996); Erenet al., Immunol.93:154-161 (1998), each of which is incorporated in its entirety by reference. Such technologies include ribosome display (Hanes et al., Proc. Natl. Acad. Sci. USA, 94:4937-4942 (Can 1997); Hanes et al., Proc. Natl. Acad. Sci. USA, 95:14130-14135 (Nov. 1998)); single-cell antibody production technology (e.g., selective lymphocyte antibody method ("SLAM") (U.S. Patent No. 5,627,052, Wen et al., "J. Immunol. 17:887-892 (1987); Babcook et al., Proc. Natl. Acad. Sci. USA 93:7843-7848 (1996)); gel microdroplet and flow cytometry (Powell et al., Biotec 8:333-337 (1990); One Cell) Examples include, but are not limited to, systems (Cambridge, MA); TM4 cells described in Gray et al., J.Imm.Meth. 182:155-163 (1995); and Kenny et al., Bio / Technol. 13:787-790 (1995); and B cell selections (Steenbakkers et al., Molec. Biol. Reports 19:125-134 (1994); and Jonak et al., Progress Biotech, Vol. 5, In Vitro Immunization in Hybridoma Technology, Borrebaeck, ed., Elsevier Science Publishers BV, Amsterdam, Netherlands (1988)).

[0056] Methods for engineering or humanizing non-human antibodies or human antibodies can also be used and are well known in the art. Generally, humanized or modified antibodies have one or more amino acid residues from non-human sources, such as, but not limited to, mouse, rat, rabbit, non-human primate, or other mammalian sources. These non-human amino acid residues are often replaced by residues called “import” residues. Such “import” residues are typically obtained from “import” variable domains, constant domains, or other domains of known human sequences.

[0057] Known human Ig sequences are disclosed, for example, at: www.ncbi.nlm.nih.gov / entrez / query.fcgi; www.ncbi.nih.gov / igblast; www.atcc.org / phage / hdb.html; www.mrc-cpe.cam.ac.uk / ALIGNMENTS.php; www.kabatdatabase.com / top.html; ftp.ncbi.nih.gov / repository / kabat; www.sciquest.com; www.abcam .com, www.antibodyresource.com / onlinecomp.html;www.public.iastate.edu / ~pedro / research_tools.html;www.whfreeman.com / immunolog y / CH05 / kuby05.htm;www.hhmi.org / grants / lectures / 1996 / vlab;www.path.cam.ac.uk / ~mrc7 / mikeimages.html;mcb.harvard.edu / BioLinks / Immunology.html;www.immunologylink.com;pathbox.wustl.edu / ~hcenter / index.html;www.appliedbiosystems.com;www.nal.usda.gov / awic / pubs / antibody;www.m.ehime-u.ac.jp / ~yasuhito / Elisa.html;www.biodesign.com;www.cancerresearchuk.org;www.biotech.ufl.edu;ww w.isac-net.org;baserv.uci.kun.nl / ~jraats / links1.html;www.recab.uni-hd.de / immuno.bme.nwu.edu;www.mrc-cpe.cam.ac.uk;www.ibt.u nam.mx / vir / V_mice.html;www.bioinf.org.uk / abs;antibody.bath.ac.uk;www.unizh.ch;www.cryst.bbk.ac.uk / ~ubcg07s;www.nimr.mrc.ac.uk / CC / ccaewg / ccaewg.html;www.path.cam.ac.uk / ~mrc7 / humanisation / TAHHP.html;www.ibt.unam.mx / vir / structure / stat_aim.html;www.biosci.missouri.edu / smithgp / index.html;www.jerini.de;Kabat et al., Sequences of Proteins of Immunological Interest, USDept.Health (1983), each is incorporated herein by reference in its entirety.

[0058] Such imported sequences can be used to reduce immunogenicity, or, as is known in the art, to reduce, enhance, or modify binding, affinity, binding rate constant, dissociation rate constant, binding activity, specificity, half-life, or any other desirable properties. Generally, CDR residues directly and almost substantially affect antigen binding. Therefore, it is also possible to replace non-human sequences in the variable and constant regions with human amino acids or other amino acids while maintaining a non-human CDR sequence or some or all of a human CDR sequence.

[0059] Antibodies may be optionally humanized, or human antibodies may be modified while retaining high affinity for antigens and other advantageous biological properties. To achieve this objective, humanized (or human) antibodies can be prepared by a process of analyzing the parent sequence and various theoretical humanized products using three-dimensional models of the parent sequence and the humanized sequence, which are optionally available. Three-dimensional immunoglobulin models are commonly available and well known to those skilled in the art. Computer programs are available that illustrate and display highly probable three-dimensional structures for selected immunoglobulin sequence candidates. By examining these displays, it is possible to analyze the likely functions of residues in the function of the immunoglobulin sequence candidate, i.e., the residues that affect the antigen-binding ability of the immunoglobulin candidate. In this way, framework (FR) residues can be selected and combined from consensus sequences and import sequences to achieve desirable antibody properties, such as enhanced affinity for target antigens.

[0060] In addition, the human anti-IL-12 / 23p40 (or anti-IL-23) specific antibody used in the method of the present invention may include a human germline light chain framework. In certain embodiments, the light chain germline sequence is selected from human VK sequences, including but not limited to A1, A10, A11, A14, A17, A18, A19, A2, A20, A23, A26, A27, A3, A30, A5, A7, B2, B3, L1, L10, L11, L12, L14, L15, L16, L18, L19, L2, L20, L22, L23, L24, L25, L4 / 18a, L5, L6, L8, L9, O1, O11, O12, O14, O18, O2, O4, and O8. For a particular purpose, this light chain human germline framework is selected from V1-11, V1-13, V1-16, V1-17, V1-18, V1-19, V1-2, V1-20, V1-22, V1-3, V1-4, V1-5, V1-7, V1-9, V2-1, V2-11, V2-13, V2-14, V2-15, V2-17, V2-19, V2-6, V2-7, V2-8, V3-2, V3-3, V3-4, V4-1, V4-2, V4-3, V4-4, V4-6, V5-1, V5-2, V5-4, and V5-6.

[0061] In other embodiments, the human anti-IL-12 / 23p40 (or anti-IL-23) specific antibody used in the method of the present invention may include a human germline heavy chain framework. In certain embodiments, this heavy chain human germline framework may include VH1-18, VH1-2, VH1-24, VH1-3, VH1-45, VH1-46, VH1-58, VH1-69, VH1-8, VH2-26, VH2-5, VH2-70, VH3-11, VH3-13, VH3-15, VH3-16, VH3-20, VH3-21, VH3-23, VH3-30, VH3- Selected from 33, VH3-35, VH3-38, VH3-43, VH3-48, VH3-49, VH3-53, VH3-64, VH3-66, VH3-7, VH3-72, VH3-73, VH3-74, VH3-9, VH4-28, VH4-31, VH4-34, VH4-39, VH4-4, VH4-59, VH4-61, VH5-51, VH6-1, and VH7-81.

[0062] In certain embodiments, the light chain variable region and / or heavy chain variable region includes a framework region, or at least a portion of the framework region (e.g., including two or three subregions such as FR2 and FR3). In certain embodiments, at least FRL1, FRL2, FRL3, or FRL4 is fully human. In other embodiments, at least FRH1, FRH2, FRH3, or FRH4 is fully human. In some embodiments, at least FRL1, FRL2, FRL3, or FRL4 is a germline sequence (e.g., human germline) or includes a human consensus sequence for a particular framework (e.g., readily available from the known human Ig sequence sources described above). In other embodiments, at least FRH1, FRH2, FRH3, or FRH4 is a germline sequence (e.g., human germline) or includes a human consensus sequence for a particular framework. In preferred embodiments, the framework region is a fully human framework region.

[0063] The humanization or engineering treatment of antibodies in the present invention is based on the works of Winter (Jone et al., Nature 321:522 (1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al., Science 239:1534 (1988)), Sims et al., J.Immunol.151:2296 (1993); Chothia and Lesk, J.Mol.Biol.196:901 (1987), Carter et al., Proc.Natl.Acad.SCi.USA89:4285 (1992); Presta et al. al., J.Immunol.151:2623 (1993), U.S. Patent No. 5723323, U.S. Patent No. 5976862, U.S. Patent No. 5824514, U.S. Patent No. 5817483, U.S. Patent No. 5814476, U.S. Patent No. 5763192, U.S. Patent No. 5723323, U.S. Patent No. 5 , 766886, 5714352, 6204023, 6180370, 5693762, 5530101, 5585089, 5225539, 4816567, International Application PCT / :US98 / 1 This can be done using any known method, including but not limited to those described in Patent No. 6280, US96 / 18978, US91 / 09630, US91 / 05939, US94 / 01234, GB89 / 01334, GB91 / 01134, GB92 / 01755, International Publication Nos. 90 / 14443, 90 / 14424, and 90 / 14430, and European Patent No. 229246 (each incorporated in its entirety by reference and including the documents cited therein).

[0064] In certain embodiments, the antibody includes a modified (e.g., a mutant) Fc region. For example, in some embodiments, the Fc region is modified to reduce or enhance the effector function of the antibody. In some embodiments, the Fc region is an isotype selected from IgM, IgA, IgG, IgE, or other isotypes. Alternatively, or in addition, it may be useful to combine amino acid modifications with one or more further amino acid modifications that modify the C1q binding and / or complement-dependent cytotoxicity function of the Fc region of the IL-23 binding molecule. A starting polypeptide for a particular purpose may be one that binds to C1q and exhibits complement-dependent cytotoxicity (CDC). Polypeptides having existing C1q binding activity and optionally the ability to further mediate CDC may be modified to enhance one or both of these activities. Amino acid modifications that modify C1q and / or modify its complement-dependent cytotoxicity function are described, for example, in International Publication No. 0042072, incorporated herein by reference.

[0065] As disclosed above, the Fc region of the human anti-IL-12 / 23p40 (or anti-IL-23) specific antibody of the present invention can be designed to have modified effector functions by, for example, modifying C1q binding and / or FcγR binding, thereby altering complement-dependent cytotoxicity (CDC) activity and / or antibody-dependent cell-mediated cytotoxicity (ADCC) activity. “Effector function” refers to a role in activating or reducing biological activity (e.g., in a target). Examples of effector functions, but not limited to, include C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, and downregulation of cell surface receptors (e.g., B cell receptors, BCRs). Such effector functions may require the Fc region to bind to a binding domain (e.g., an antibody variable domain) and can be evaluated using a wide variety of assays (e.g., Fc binding assays, ADCC assays, CDC assays, etc.).

[0066] For example, a variant Fc region of a human anti-IL-12 / 23p40 (or anti-IL-23) antibody can be generated that has improved C1q binding and improved FcγRIII binding (e.g., having both improved ADCC activity and improved CDC activity). Alternatively, if it is desired to reduce or eliminate effector function, the variant Fc region can be modified to reduce CDC activity and / or ADCC activity. In other embodiments, only one of these activities may be enhanced, and the other activity may be selectively reduced at the same time (e.g., to generate an Fc region variant having improved ADCC activity and reduced CDC activity, and the reverse Fc region variant).

[0067] Fc mutations can also be introduced by genetically engineering genes to alter their interaction with the embryonic Fc receptor (FcRn) and improve their pharmacokinetic properties. The collection of human Fc variants with improved binding to FcRn has been described (Shields et al., (2001). High resolution mapping of the binding site on human IgG1 for FcγRI, FcγRII, FcγRIII, and FcRn and design of IgG1 variants with improved binding to the FCγR, J. Biol. Chem. 276:6591-6604).

[0068] Other types of amino acid substitutions can help alter the glycosylation pattern of the Fc region of human anti-IL-12 / 23p40 (or anti-IL-23) specific antibodies. Glycosylation of the Fc region is typically either N-linked or O-linked. N-linked glycosylation refers to the attachment of a carbohydrate moiety to the side chain of an asparagine residue. O-linked glycosylation refers to the attachment of one of the following sugars to a hydroxyamino acid, most commonly serine or threonine: N-acetylgalactosamine, galactose, or xylose, although 5-hydroxyproline or 5-hydroxylysine may also be used. The recognition sequences for the enzymatic attachment of the carbohydrate moiety to the asparagine side-chain peptide sequence are asparagine-X-serine and asparagine-X-threonine, where X is any amino acid other than proline. Therefore, the presence of either of these peptide sequences in a polypeptide results in a potential glycosylation site.

[0069] The glycosylation pattern can be altered, for example, by deleting one or more glycosylation sites found in the polypeptide, and / or by adding one or more glycosylation sites that are not present in the polypeptide. Addition of glycosylation sites to the Fc region of human IL-23-specific antibodies is successfully achieved by modifying the amino acid sequence to include one or more of the tripeptide sequences described above (in the case of N-linked glycosylation sites). An exemplary glycosylation variant has an amino acid substitution at the heavy chain residue Asn297. This modification can also be achieved by adding or substituting one or more serine or threonine residues into the original polypeptide sequence (in the case of O-linked glycosylation sites). Additionally, changing Asn 297 to Ala can remove one of the glycosylation sites.

[0070] In certain embodiments, the human anti-IL-12 / 23p40 (or anti-IL-23) specific antibody of the present invention is expressed in cells expressing beta(1,4)-N-acetylglucosaminyltransferase III (GnT III) such that GnT III adds GlcNAc to the human anti-IL-12 / 23p40 (or anti-IL-23) antibody. Methods for producing the antibody in this manner are provided in International Publication No. 9954342, No. 03011878, Japanese Patent Publication No. 20030003097(A1), and Umana et al., Nature Biotechnology, 17:176-180, Feb. 1999, all of which are specifically incorporated herein by reference in their entirety.

[0071] Human anti-IL-12 / 23p40 (or anti-IL-23) antibodies can also be optionally produced by immunization of transgenic animals (e.g., mice, rats, hamsters, non-human primates, etc.) capable of producing a repertoire of human antibodies, as described herein and / or known in the art. Cells producing human anti-IL-12 / 23p40 (or anti-IL-23) antibodies may be isolated from such animals and immortalized using preferred methods, such as those described herein.

[0072] Transgenic mice capable of producing a repertoire of human antibodies that bind to human antigens can be produced by known methods (e.g., but not limited to these, U.S. Patents No. 5,770,428, 5,569,825, 5,545,806, 5,625,126, 5,625,825, 5,633,425, 5,661,016, and 5,789 issued to Lonberg et al.) , Patent No. 650, International Publication No. 98 / 50433 by Jakobovits et al., International Publication No. 98 / 24893 by Jakobovits et al., International Publication No. 98 / 24884 by Lonberg et al., International Publication No. 97 / 13852 by Lonberg et al., International Publication No. 94 / 25585 by Lonberg et al., International Publication No. 96 / 34096 by Kucherlapate et al., European Patent No. 0463 by Kucherlapate et al. See also: Patent No. 151(B1), European Patent No. 0710 719(A1) by Kucherlapate et al., U.S. Patent No. 5,545,807 by Surani et al., International Publication No. 90 / 04036 by Bruggemann et al., European Patent No. 0438 474(B1) by Bruggemann et al., European Patent No. 0814 259(A2) by Lonberg et al., UK Patent No. 2 272 440(A) by Lonberg et al., Nature 368 856-859 (1994) by Lonberg et al., Int.Immunol. 6(4) 579-591 (1994) by Taylor et al., Nature Genetics 7:13-21 (1994) by Green et al., and Nature Genetics by Mendez et al. These can be produced by 15:146-156 (1997), Taylor et al. Nucleic Acids Research 20(23):6287-6295 (1992), Tuaillon et al. Proc Natl Acad Sci USA 90(8)3720-3724 (1993), Lonberg et al. Int Rev Immunol 13(1):65-93 (1995), and Fishwald et al. Nat Biotechnol 14(7):845-851 (1996), each of which is incorporated herein by reference in its entirety.Generally, these mice contain at least one transgene containing DNA derived from at least one human immunoglobulin locus that is functionally reconstituted or capable of undergoing functional reconstitution. By disrupting or deleting the endogenous immunoglobulin locus in such mice, the mice can be deprived of the ability to produce antibodies encoded by the endogenous gene.

[0073] Screening of antibodies for specific binding to similar proteins or fragments can be successfully achieved using peptide display libraries. This method involves screening a large collection of peptides for individual members with desired function or structure. Antibody screening of peptide display libraries is well known in the art. The length of the displayed peptide sequences is 3 to 5000 or more amino acids, frequently 5 to 100 amino acids, and often about 8 to 25 amino acids. In addition to direct chemical synthesis methods for creating peptide libraries, several recombinant DNA methods have also been described. One type involves displaying peptide sequences on the surface of bacteriophages or cells. Each bacteriophage or cell contains a nucleotide sequence encoding a specific displayed peptide sequence. Such methods are described in International Patent Publications 91 / 17271, 91 / 18980, 91 / 19818, and 93 / 08278.

[0074] Other systems for preparing peptide libraries include both in vitro chemical synthesis and recombinant methods. See International Patent Publications 92 / 05258, 92 / 14843, and 96 / 19256. See also U.S. Patents 5,658,754 and 5,643,768. Peptide display libraries, vectors, and screening kits are commercially available from suppliers such as Invitrogen (Carlsbad, CA) and Cambridge Antibody Technologies (Cambridgeshire, UK). For example, U.S. patent numbers 4,704692, 4,939666, 4,946778, 5260203, 5455030, 5518889, 5534621, 5656730, 5763733, 5767260, and 5856456, which were transferred to Enzon; ​​U.S. patent numbers 5,223409, 5403484, 5571698, and 5837500, which were transferred to Dyax; and U.S. patent numbers 5,427908 and 5580717, which were transferred to Affymax, as well as Cambridge antibody. See U.S. Patent No. 5,885,793, assigned to Technologies; U.S. Patent No. 5,750,373, assigned to Genentech; U.S. Patents No. 5,618,920, No. 5,595,898, No. 5,576,195, No. 5,698,435, No. 5,693,493, and No. 5,698,417, assigned to Xoma, Colligan, Ausubel, or Sambrook, each of which is incorporated herein by reference in its entirety.

[0075] The antibodies used in the methods of the present invention may also be prepared using at least one anti-IL-12 / 23p40 (or anti-IL-23) antibody encoding a nucleic acid to provide transgenic animals or mammals such as goats, cattle, horses, sheep, and rabbits that produce such antibodies in their milk. Such animals can be prepared using known methods. See, for example, but not limited to, U.S. Patents 5,827,690, 5849992, 4873316, 5849992, 5994616, 5565362, 5,304,489, and others, each of which is incorporated herein by reference in whole.

[0076] The antibodies used in the methods of the present invention can be further prepared using at least one anti-IL-12 / 23p40 (or anti-IL-23) antibody encoding a nucleic acid to provide transgenic plants and cultured plant cells (e.g., tobacco and maize) that produce such antibodies, specified parts, or mutants in plant parts or cells cultured therefrom. As a non-limiting example, large quantities of recombinant proteins have been successfully provided using, for example, transgenic tobacco leaves expressing recombinant proteins with an inducible promoter. See, for example, Cramer et al., Curr. Top. Microbol. Immunol. 240:95-118 (1999) and the literature cited therein. Transgenic maize has also been used to express mammalian proteins with biological activity equivalent to proteins produced in other recombinant systems or purified from natural resources at a commercially viable level. See, for example, Hood et al., Adv. Exp. Med. Biol. 464:127-147 (1999) and the literature cited therein. Antibodies, including antibody fragments such as single-chain antibodies (scFv), have also been produced in large quantities from the seeds of transgenic plants such as tobacco seeds and potato tubers. See, for example, Conradr, Plant Mol. Biol. 38:101-109 (1998) and the literature cited therein. Therefore, the antibodies of the present invention can also be produced using transgenic plants according to known methods. See, for example, Fischer et al., Biotechnol. Appl. Biochem. 30:99-108 (Oct., 1999), Ma et al., Trends Biotechnol. 13:522-7 (1995); Ma et al., Plant Physiol. 109:341-6 (1995); Whitelam et al., Biochem. Soc. Trans. 22:940-944 (1994) and the literature cited therein. Each of the above references is incorporated herein by reference in its entirety.

[0077] The antibodies used in the method of the present invention can bind to human IL-12 / IL-23p40 or IL-23 with a wide range of affinity (KD). In preferred embodiments, human mAbs can optionally bind to human IL-12 / IL-23p40 or IL-23 with high affinity. For example, human mAbs can bind to human IL-12 / IL-23p40 or IL-23 with a KD of about 10⁻⁷ M or less, for example, but not limited to, 0.1 to 9.9 (or any range or value therein) x 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹¹, 10⁻¹², 10⁻¹³, ​​or any range or value therein.

[0078] The affinity or binding activity of an antibody to an antigen can be determined experimentally using any suitable method. (See, for example, Berzofsky, et al., "Antibody-Antigen Interactions," In Fundamental Immunology, Paul, WE, Ed., Raven Press: New York, NY (1984); Kuby, Janis Immunology, WH Freeman and Company: New York, NY (1992); and the methods described herein). The affinity measured for a particular antibody-antigen interaction may differ when measured under different conditions (e.g., salt concentration, pH). Therefore, the measurement of affinity and other antigen-binding parameters (e.g., KD, Ka, Kd) is preferably performed using standardized solutions of the antibody and antigen, and standardized buffers such as the buffers described herein.

[0079] Vectors and host cells The present invention also relates to a vector comprising an isolated nucleic acid molecule, a host cell genetically engineered with a recombinant vector, and the production of at least one anti-IL-12 / IL-23p40 antibody by recombinant techniques well known in the art. See, for example, Sambrook et al. and Ausubel et al. above, each of which is incorporated herein by reference in whole.

[0080] Polynucleotides can be selectively bound to vectors containing selection markers for host proliferation. Generally, plasmid vectors are introduced into precipitates such as calcium phosphate precipitates or into complexes with charged lipids. If the vector is a virus, it can be packaged in vitro using a suitable packaging cell line and then transduced into host cells.

[0081] The DNA insert should be functionally linked to an appropriate promoter. The expression construct further includes a transcription start site, a transcription termination site, and, within the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcript expressed by the construct preferably includes translation beginning with start and stop codons (e.g., UAA, UGA, or UAG) appropriately located at the end of the mRNA to be translated, with UAA and UAG being preferred for expression in mammalian or eukaryotic cells.

[0082] The expression vector preferably contains at least one selection marker, but this is arbitrary selection. Such markers include, for example, methotrexate (MTX) for eukaryotic cell culture and dihydrofolate reductase (dihydrofolate). This includes, but is not limited to, genes for resistance to reductase, DHFR, U.S. Patent Nos. 4,399,216, 4,634,665, 4,656,134, 4,956,288, 5,149,636, and 5,179,017, ampicillin, neomycin (G418), mycophenolic acid, or glutamine synthetase (GS, U.S. Patents Nos. 5,122,464, 5,770,359, and 5,827,739), and tetracycline or ampicillin resistance genes for culture in Escherichia coli (E. coli) and other bacteria or prokaryotes (the above patents are incorporated herein by reference in their entirety). Appropriate culture media and conditions for the above-mentioned host cells are known in the art. Suitable vectors will be readily apparent to those skilled in the art. Introduction of the vector construct into host cells can be achieved by calcium phosphate transfection, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, or other known methods. Such methods are described in the art above, such as in Sambrook, Chapters 1-4 and 16-18, and Ausubel, Chapters 1, 9, 13, 15, and 16.

[0083] At least one antibody used in the method of the present invention may be expressed in a modified form, such as a fusion protein, and may include not only secretory signals but also additional heterologous functional regions. For example, a region of additional amino acids, particularly charged amino acids, can be added to the N-terminus of the antibody to improve stability and persistence in host cells during purification or subsequent processing and storage. Alternatively, a peptide portion can be added to the antibody of the present invention to facilitate purification. Such regions can be removed before the final preparation of the antibody or at least one fragment thereof. Such methods are described in many standard laboratory manuals, such as Sambrook, Chapters 17.29-17.42 and 18.1-18.74, and Ausubel, Chapters 16, 17 and 18.

[0084] Those skilled in the art will be familiar with the numerous expression systems available for expressing the nucleic acids encoding the proteins used in the methods of the present invention. Alternatively, the nucleic acids can be expressed in host cells containing endogenous DNA encoding antibodies by being (operationally) switched on. Such methods are well known in the art, as described in U.S. Patents 5,580,734, 5,641,670, 5,733,746, and 5,733,761, which are incorporated herein by reference in their entirety.

[0085] Mammalian cells are an example of a cell culture useful for producing antibodies, their identified parts, or variants. Mammalian cell lines often take the form of a single layer of cells, but suspensions or bioreactors of mammalian cells can also be used. Several suitable host cell lines capable of expressing intact glycosylated proteins have been developed in the art, including COS-1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-1651), HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e.g., ATCC CRL1610), and BSC-1 (e.g., ATCC CRL-26) cell lines, Cos-7 cells, CHO cells, hep G2 cells, P3X63Ag8.653, SP2 / 0-Ag14, 293 cells, and HeLa cells, which are readily available, for example, from the American Type Culture Collection (Manassas, Va) (www.atcc.org). Preferred host cells include lymphoid cells such as myeloma and lymphoma cells. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC deposit number CRL-1580) and SP2 / 0-Ag14 cells (ATCC deposit number CRL-1851). In a particularly preferred embodiment, the recombinant cells are P3X63Ab8.653 or SP2 / 0-Ag14 cells.

[0086] These cell expression vectors may contain one or more regulatory sequences, including, but not limited to, origins of replication, promoters (e.g., late or early SV40 promoter, CMV promoter (U.S. Patent No. 5,168,062, 5,385,839), HSV tk promoter, pgk (phosphoglycerate kinase) promoter, EF-1α promoter (U.S. Patent No. 5,266,491), at least one human immunoglobulin promoter, enhancer, and / or ribosome binding site, RNA splice site, polyadenylation site (e.g., SV40 large T Ag polyaddition site), and processing information sites such as transcription termination sequences. See, for example, Ausubel et al. and Sambrook et al. above. Other cells useful for generating nucleic acids or proteins of the present invention are known and / or available from, for example, the American Type Culture Collection Catalogue of Cell Lines and Hybridomas (www.atcc.org) or other known or commercial sources.

[0087] When eukaryotic host cells are used, typically, polyadenylation or transcription termination sequences are incorporated into the vector. An example of a termination sequence is a polyadenylation sequence from the bovine growth hormone gene. Sequences for precise transcriptional splicing can also be included. An example of a splicing sequence is the VP1 intron from SV40 (Sprague, et al., J. Virol. 45:773-781 (1983)). In addition, as is known in the art, gene sequences for controlling replication within the host cell can be incorporated into the vector.

[0088] Antibody purification Anti-IL-12 / IL-23p40 or IL-23 antibodies can be recovered and purified from recombinant cell cultures by known methods, including but not limited to protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxyl apatite chromatography, and lectin chromatography. High-performance liquid chromatography (HPLC) can also be used for purification. For example, see Chapters 1, 4, 6, 8, 9, and 10 of Colligan, Current Protocols in Immunology or Current Protocols in Protein Science, John Wiley & Sons, NY, NY (1997-2001), each incorporated herein by reference.

[0089] The antibodies used in the methods of the present invention include naturally purified products, products from chemical synthesis processes, and products produced by recombinant technology from eukaryotic hosts, including, for example, yeast, higher plants, insects, and mammalian cells. Depending on the host used in the recombinant production process, the antibodies may be glycosylated or not, but glycosylation is preferred. Such methods are described in many standard laboratory manuals, such as Sambrook, sections 17.37–17.42, Ausubel, chapters 10, 12, 13, 16, 18, and 20, and Colligan, Protein Science, chapters 12–14, all of which are incorporated herein by reference in their entirety.

[0090] Anti-IL-12 / IL-23p40 or IL-23 antibody The anti-IL-12 / IL-23p40 or IL-23 antibody according to the present invention comprises any protein or peptide-containing molecule that can be incorporated into the antibody, including, for example, at least one ligand-binding portion (LBP), for example, a complementarity-determining region (CDR) of the heavy or light chain or its ligand-binding portion, a variable region of the heavy or light chain, a framework region (e.g., FR1, FR2, FR3, FR4, or fragments thereof, and optionally including at least one substitution, insertion, or deletion), a constant region of the heavy or light chain (e.g., at least one CH1, hinge 1, hinge 2, hinge 3, hinge 4, CH2, or CH3, or fragments thereof, and optionally including at least one substitution, insertion, or deletion), or any portion thereof. Antibodies may include, or be derived from, any mammal, including but not limited to, humans, mice, rabbits, rats, rodents, primates, or any combination thereof.

[0091] Preferably, a human antibody or antigen-binding fragment binds to human IL-12 / IL-23p40 or IL-23, thereby partially or substantially neutralizing at least one biological activity of the protein. An antibody or identified portion or variant thereof that partially or preferably substantially neutralizes at least one biological activity of at least one IL-12 / IL-23p40 or IL-23 protein or fragment can bind to the protein or fragment, thereby inhibiting the activity of IL-12 / IL-23p40 or IL-23 mediated by binding to IL-12 and / or IL-23 receptors, or by other IL-12 / IL-23p40 or IL-23-dependent or mediated mechanisms. As used herein, the term “neutralizing antibody” refers to an antibody capable of inhibiting IL-12 / IL-23p40 or IL-23-dependent activity by about 20–120%, preferably at least about 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% or more, depending on the assay. The ability of an anti-IL-12 / IL-23p40 or IL-23 antibody to inhibit IL-12 / IL-23p40 or IL-23-dependent activity is preferably evaluated by at least one suitable IL-12 / IL-23p40 or IL-23 protein or receptor assay described herein and / or known in the art. Human antibodies may be of any class (IgG, IgA, IgM, IgE, IgD, etc.) or isotype, and may include kappa or lambda light chains. In one embodiment, the human antibody comprises an IgG heavy chain or a defined fragment, e.g., at least one isotype from IgG1, IgG2, IgG3, or IgG4 (e.g., γ1, γ2, γ3, γ4). This type of antibody can be prepared by utilizing a transgenic mouse or other non-human transgenic mammal containing at least one human light chain (e.g., IgG, IgA, and IgM) transgene described herein and / or known in the art. In another embodiment, the anti-IL-23 human antibody comprises an IgG1 heavy chain and an IgG1 light chain.

[0092] The antibody binds to at least one identified epitope specific to at least one IL-12 / IL-23p40 or IL-23 protein, subunit, fragment, portion, or any combination thereof. This at least one epitope may include at least one antibody-binding region comprising at least a portion of the protein, and this epitope preferably consists of at least one extracellular portion, soluble portion, hydrophilic portion, outer portion, or cytoplasmic portion of the protein.

[0093] Generally, a human antibody or antigen-binding fragment comprises an antigen-binding region comprising at least one human complementarity-determining region (CDR1, CDR2, and CDR3) or at least one heavy chain variable region variant, and at least one human complementarity-determining region (CDR1, CDR2, and CDR3) or at least one light chain variable region variant. The CDR sequences may be derived from human germline sequences or may be strictly identical to germline sequences. For example, CDRs from a synthetic library derived from original non-human CDRs can be used. These CDRs may be formed by incorporating conserved substitutions derived from the original non-human sequences. In another particular embodiment, the antibody or antigen-binding moiety or variant may have an antigen-binding region comprising at least a portion of at least one light chain CDR (i.e., CDR1, CDR2, and / or CDR3) having the corresponding CDR1, 2, and / or 3 amino acid sequences.

[0094] Such antibodies can be prepared by chemically linking various parts of an antibody (e.g., CDR, framework) together using conventional techniques, either by preparing and expressing nucleic acid molecules encoding the antibody (i.e., one or more) using conventional techniques relating to recombinant DNA technology, or by using any other suitable method.

[0095] In one embodiment, the anti-IL-12 / 23p40 antibody useful for the present invention is a monoclonal antibody, preferably a human mAb, comprising the heavy chain complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3 of SEQ ID NOs. 1, 2, and 3, respectively, and the light chain CDRs LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 4, 5, and 6, respectively.

[0096] An anti-IL-12 / IL-23p40 or IL-23 specific antibody may include at least one of a heavy chain or light chain variable region having a defined amino acid sequence. For example, in a preferred embodiment, the anti-IL-12 / IL-23p40 or IL-23 antibody includes an anti-IL-12 / IL-23p40 antibody having a heavy chain variable region containing at least 85%, preferably at least 90%, more preferably at least 95%, and most preferably 100%, the same amino acid sequence as SEQ ID NO: 7, and a light chain variable region containing at least 85%, preferably at least 90%, more preferably at least 95%, and most preferably 100%, the same amino acid sequence as SEQ ID NO: 8.

[0097] An anti-IL-12 / IL-23p40 or IL-23 specific antibody may also include at least one of a heavy chain or a light chain having a defined amino acid sequence. In another preferred embodiment, the anti-IL-12 / IL-23p40 or IL-23 antibody comprises an anti-IL-12 / IL-23p40 antibody having a heavy chain containing at least 85%, preferably at least 90%, more preferably at least 95%, and most preferably 100% identical amino acid sequence in SEQ ID NO: 10, and a light chain variable region containing at least 85%, preferably at least 90%, more preferably at least 95%, and most preferably 100% identical amino acid sequence in SEQ ID NO: 11.

[0098] Preferably, the anti-IL-12 / 23p40 antibody is ustekinumab (Stelara®) that includes a heavy chain having the amino acid sequence of SEQ ID NO: 10 and a light chain having the amino acid sequence of SEQ ID NO: 11. Other examples of anti-IL12 / 23p40 antibodies useful in the present invention include briakinumab (ABT-874, Abbott) and other antibodies described in U.S. Patent Nos. 6,914,128, 7,247,711, 7,700,739 (the entire contents of which are incorporated herein by reference), but are not limited thereto.

[0099] The present invention also relates to antibodies, antigen-binding fragments, immunoglobulin chains, and CDRs that include amino acids in sequences that are substantially the same as the amino acid sequences described herein. Preferably, such antibodies or antigen-binding fragments and antibodies that include such chains or CDRs have high affinity (e.g., KD is about 10 -9 M or less) and can bind to human IL-12 / IL-23p40 or IL-23. Amino acid sequences that are substantially the same as the sequences described herein include sequences that include conservative amino acid substitutions and amino acid deletions and / or insertions. Conservative amino acid substitutions refer to substituting a first amino acid with a second amino acid that has similar chemical and / or physical properties (e.g., charge, structure, polarity, hydrophobicity / hydrophilicity) to those of the first amino acid. Conservative substitutions include, but are not limited to, substituting one amino acid with another amino acid within the following groups: lysine (K), arginine (R), and histidine (H); aspartate (D) and glutamate (E); asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine (C), and glycine (G); F, W, and Y; C, S, and T.

[0100] Antibodies conjugating to human IL-12 / IL-23p40 or IL-23 and containing a defined heavy chain or light chain variable region can be prepared using preferred methods, such as phage display (Katsube, Y., et al., Int J Mol. Med, 1(5):863-868 (1998)) or methods employing transgenic animals, as known in the art and / or described herein. For example, a transgenic mouse containing a functionally reconstituted human immunoglobulin heavy chain transgene and a transgene containing DNA from a human immunoglobulin light chain locus capable of functional reconstitution can be immunized with human IL-12 / IL-23p40 or IL-23 or a fragment thereof to induce antibody production. If desired, antibody-producing cells can be isolated, and hybridomas or other immortalized antibody-producing cells can be prepared as described herein and / or as known in the art. Alternatively, the antibody, specified region, or variant can be expressed in a suitable host cell using the encoding nucleic acid or a portion thereof.

[0101] The anti-IL-12 / IL-23p40 or IL-23 antibody used in the method of the present invention may include substitutions, deletions, or additions of one or more amino acids, either by spontaneous mutation or human manipulation, as specified herein.

[0102] The number of amino acid substitutions that a person skilled in the art can perform depends on many factors, including those mentioned above. Generally speaking, the number of amino acid substitutions, insertions, or deletions in a given anti-IL-12 / IL-23p40 or IL-23 antibody, fragment, or variant shall not exceed 40, 30, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, for example, 1 to 30 or any range or value within this range, as specified herein.

[0103] The functionally essential amino acids within anti-IL-12 / IL-23p40 or IL-23 specific antibodies can be identified by methods known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (e.g., Ausubel, Chapters 8, 15; Cunningham and Wells, Science 244:1081-1085 (1989)). In the latter procedure, a single alanine mutation is introduced at each residue within the molecule. The resulting mutant molecules are then tested for biological activity, such as, but not limited to, at least one IL-12 / IL-23p40 or IL-23 neutralizing activity. Sites that are extremely important for antibody binding can also be identified by structural analysis such as crystallization, nuclear magnetic resonance, or photoaffinity labeling (Smith et al., J.Mol.Biol.224:899-904 (1992) and de Vos et al., Science 255:306-312 (1992)).

[0104] Anti-IL-12 / IL-23p40 or IL-23 antibodies may include, but are not limited to, at least one moiety, sequence, or combination selected from all five of the adjacent amino acids of at least one of the sequences in SEQ ID NOs. 1, 2, 3, 4, 5, 6, 7, 8, 10, or 11.

[0105] IL-12 / IL-23p40 or IL-23 antibody or specific part or variant may include, but is not limited to, at least one part, sequence, or combination selected from at least 3 to 5 adjacent amino acids of the above SEQ ID NO:, 5 to 17 adjacent amino acids of the above SEQ ID NO:, 5 to 10 adjacent amino acids of the above SEQ ID NO:, 5 to 11 adjacent amino acids of the above SEQ ID NO:, 5 to 7 adjacent amino acids of the above SEQ ID NO:, or 5 to 9 adjacent amino acids of the above SEQ ID NO:.

[0106] The anti-IL-12 / IL-23p40 or IL-23 antibody may further optionally contain at least one polypeptide comprising 70-100% of the adjacent amino acids of sequence numbers 5, 17, 10, 11, 7, 9, 119, 108, 449, or 214. In one embodiment, the amino acid sequence of an immunoglobulin chain, or a portion thereof (e.g., a variable region, CDR), has about 70-100% identity with the amino acid sequence of at least one corresponding chain from the sequence numbers (e.g., 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or any range or value within this). For example, the amino acid sequence of the light chain variable region can be compared with the sequence of the above sequence number, or the amino acid sequence of the heavy chain CDR3 can be compared with the above sequence number. Preferably, 70-100% amino acid identity (i.e., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or any range or value therein) is determined using a suitable computer algorithm, as is known in the art.

[0107] As is known in the art, "identity" is the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, determined by comparing their sequences. In the art, "identity" also means the degree of sequence relevance between polypeptide or polynucleotide sequences, as determined by the correspondence between such linear sequences. "Identity" and "similarity" can be easily calculated by known methods, including, but are not limited to, those described in Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., Siam J. Applied Math., 48:1073 (1988). In addition, values ​​related to the degree of identity can be obtained from amino acid and nucleotide sequence alignments created using the default settings of AlignX, a component of Vector NTI Suite 8.0 (Informax, Frederick, MD).

[0108] A preferred method for determining identity is designed to obtain the greatest degree of agreement between the sequences being tested. Methods for determining identity and similarity are codified in publicly available computer programs. Preferred computer program methods for determining identity and similarity between two sequences include, but are not limited to, the GCG program package (Devereux, J, et al., Nucleic Acids Research 12(1):387 (1984)), BLASTP, BLASTN, and FASTA (Atschul, SF et al., J. Molec. Biol. 215:403-410 (1990)). The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBINLM NIH Bethesda, Md.20894: Altschul, S., et al., J.NIHMol.Biol.215:403-410 (1990)). The well-known Smith-Waterman algorithm can also be used to determine identity.

[0109] Exemplary heavy chain and light chain variable region sequences, and portions thereof, are shown in the above sequence numbers. The antibody of the present invention, or a specified variant thereof, may contain any number of adjacent amino acid residues from the antibody of the present invention, the number of which is selected from an integer group consisting of 10 to 100% of the number of adjacent residues in the anti-IL-12 / IL-23p40 or IL-23 antibody. Optionally, this subsequence of adjacent amino acids may have an amino acid length of at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250 or more, or any range or value therein. Furthermore, the number of such subsequences may be any integer selected from the group consisting of 1 to 20, such as at least 2, 3, 4, or 5.

[0110] As will be apparent to those skilled in the art, the present invention comprises at least one biologically active antibody of the present invention. The biologically active antibody has a specific activity of at least 20%, 30%, or 40%, preferably at least 50%, 60%, or 70%, most preferably at least 80%, 90%, or 95% to 100% or more (but not limited to, up to 10 times the specific activity) of natural (non-synthetic), endogenous, or related and known antibodies. Methods for assaying and quantitatively measuring enzyme activity and substrate specificity are well known to those skilled in the art.

[0111] In another embodiment, the present invention relates to human antibodies and antigen-binding fragments described herein that are modified by covalent bonding of an organic moiety. Such modifications can produce antibodies or antigen-binding fragments with improved pharmacokinetic properties (e.g., increased in vivo serum half-life). The organic moiety may be a linear or branched hydrophilic polymer group, a fatty acid group, or a fatty acid ester group. In certain embodiments, the hydrophilic polymer group may be a polyalkane glycol (e.g., polyethylene glycol (PEG), polypropylene glycol (PPG)), a carbohydrate polymer, an amino acid polymer, or polyvinylpyrrolidone, with a molecular weight of about 800 to about 120,000 daltons, and the fatty acid group or fatty acid ester group may contain about 8 to about 40 carbon atoms.

[0112] Modified antibodies and antigen-binding fragments may contain one or more organic moieties that are directly or indirectly covalently bound to the antibody. Each organic moiety bound to the antibody or antigen-binding fragment of the present invention may independently be a hydrophilic polymer group, a fatty acid group, or a fatty acid ester group. As used herein, the term “fatty acid” includes monocarboxylic acids and dicarboxylic acids. As used herein, the term “hydrophilic polymer group” means an organic polymer that is more soluble in water than octane. For example, polylysine is more soluble in water than octane. Therefore, antibodies modified by covalent bonding of polylysine are included in the present invention. Suitable hydrophilic polymers for modifying the antibodies of the present invention may be linear or branched and include, for example, polyalkane glycols (e.g., PEG, monomethoxy-polyethylene glycol (mPEG), PPG, etc.), carbohydrates (e.g., dextran, cellulose, oligosaccharides, polysaccharides, etc.), polymers of hydrophilic amino acids (e.g., polylysine, polyarginine, polyaspartic acid, etc.), polyalkane oxides (e.g., polyethylene oxide, polypropylene oxide, etc.), and polyvinylpyrrolidone. Preferably, the hydrophilic polymers for modifying the antibodies of the present invention have a molecular weight of about 800 to about 150,000 daltons as individual molecules. For example, PEG5000 and PEG20,000 can be used. The subscript indicates the average molecular weight (daltons) of the polymer. The hydrophilic polymer groups can be substituted with 1 to about 6 alkyl groups, fatty acid groups, or fatty acid ester groups. Hydrophilic polymers substituted with fatty acid or fatty acid ester groups can be prepared by using suitable methods. For example, a polymer containing an amine group can be linked to a carboxylate salt of a fatty acid or fatty acid ester, and an activated carboxylate salt on a fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer.

[0113] Fatty acids and fatty acid esters suitable for modifying the antibodies of the present invention may be saturated or may contain one or more unsaturated units. Examples of fatty acids suitable for modifying the antibodies of the present invention include n-dodecanoic acid (C12, lauric acid), n-tetradecanoic acid (C14, myristic acid), n-octadecanoic acid (C18, stearic acid), n-eicosanoic acid (C20, arachidic acid), n-docosanoic acid (C22, behenic acid), n-triacontanoic acid (C30), n-tetracontanoic acid (C40), cis-Δ9-octadecanoic acid (C18, oleic acid), all cis-Δ5,8,11,14-eicosatetraenoic acid (C20, arachidonic acid), octanedionic acid, tetradecanedionic acid, octadecanedionic acid, and docosanedionic acid. Suitable fatty acid esters include monoesters of dicarboxylic acids containing a linear or branched lower alkyl group. The lower alkyl group may contain 1 to about 12 carbon atoms, preferably 1 to about 6 carbon atoms.

[0114] Modified human antibodies and antigen-binding fragments can be prepared using preferred methods, such as reacting them with one or more modifying agents. As used herein, the term “modifying agent” means a preferred organic group containing an activating group (e.g., hydrophilic polymers, fatty acids, fatty acid esters). An “activating group” is a chemical moiety or functional group that reacts with a second chemical group under appropriate conditions, thereby forming a covalent bond between the modifying agent and the second chemical group. Examples of amine-reactive activating groups include tosylic acid, mesylic acid, electrophilic groups such as halo(chloro, bromo, fluoro, iodine), and N-hydroxysuccinimidyl esters (NHS). Examples of activating groups that can react with thiols include maleimide, iodoacetyl, acryloryl, pyridyl disulfide, and 5-thiol-2-nitrobenzoic acid thiol (TNB-thiol). Aldehyde functional groups can be linked to amine- or hydrazide-containing molecules, and azide groups can react with trivalent phosphorus groups to form phosphoramidate or phosphorimide bonds. Preferred methods for introducing activating groups into molecules are known in the art (see, for example, Hermanson, GT, Bioconjugate Techniques, Academic Press: San Diego, CA (1996)). Activating groups can be linked directly to organic groups (e.g., hydrophilic polymers, fatty acids, fatty acid esters) or via linker moieties (e.g., divalent C1-C12 groups, where one or more carbon atoms can be substituted with heteroatoms such as oxygen, nitrogen, or sulfur). Suitable linker moieties include, for example, tetraethylene glycol, -(CH2)3-, -NH-(CH2)6-NH-, -(CH2)2-NH-, and -CH2-O-CH2-CH2-O-CH2-CH2-O-CH-NH-.Modifiers containing a linker moiety can be produced by reacting a mono-Boc-alkyldiamine (e.g., mono-Boc-ethylenediamine, mono-Boc-diaminohexane) with a fatty acid in the presence of, for example, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), thereby forming an amide bond between the free amine and the fatty acid carboxylate. The Boc protecting group can be removed from the product by treatment with trifluoroacetic acid (TFA) to expose a primary amine that can be linked to another carboxylate as described, or it can be reacted with maleic anhydride to cyclize the resulting product to produce an activated maleimide derivative of the fatty acid. (See, for example, International Publication No. 92 / 16221 (Thompson et al.), by which this entire teaching is incorporated herein by reference).

[0115] Modified antibodies can be produced by reacting human antibodies or antigen-binding fragments with a modifying agent. For example, the organic portion can be conjugated to the antibody in a non-site-specific manner using an amine-reactive modifier, such as the NHS ester of PEG. Modified human antibodies or antigen-binding fragments can also be prepared by reducing the disulfide bonds (e.g., intrachain disulfide bonds) of the antibody or antigen-binding fragment. In this case, the modified antibody of the present invention can be produced by reacting the reduced antibody or antigen-binding fragment with a thiol-reactive modifier. Modified human antibodies and antigen-binding fragments containing organic moieties that bind to specific sites on the antibodies of the present invention can be prepared using appropriate methods such as reverse proteolysis (Fisch et al., Bioconjugate Chem., 3:147-153 (1992), Werlen et al., Bioconjugate Chem., 5:411-417 (1994), Kumaran et al., Protein Sci. 6(10):2233-2241 (1997); Itoh et al., Bioorg. Chem., 24(1):59-68 (1996), Capellas et al., Biotechnol. Bioeng., 56(4):456-463 (1997)), and methods described in Hermanson, GT, Bioconjugate Techniques, Academic Press: San Diego, CA (1996).

[0116] The method of the present invention also uses an anti-IL-12 / IL-23p40 or IL-23 antibody composition comprising at least one, at least two, at least three, at least four, at least five, at least six or more of its anti-IL-12 / IL-23p40 or IL-23 antibodies, provided in non-spontaneous compositions, mixtures, or forms as described herein and / or known in the art. Such a composition comprises a non-spontaneous composition comprising at least one or two full-length, C and / or N-terminal deletion variants, domains, fragments, or identified variants of the amino acid sequence of the anti-IL-12 / IL-23p40 or IL-23 antibody, selected from the group consisting of 70-100% of the adjacent amino acids of the above SEQ ID NO: or identified fragments, domains, or variants thereof. A preferred anti-IL-12 / IL-23p40 or IL-23 antibody composition includes, for example, 70 to 100% of the above SEQ ID NO: 70 to 100% of the anti-IL-12 / IL-23p40 or IL-23 antibody sequence described herein, or a specified fragment, domain, or variant thereof, as at least one CDR or LBP-containing portion, and at least one full-length fragment, domain, or variant. A more preferred composition includes, for example, 70 to 100% of the above SEQ ID NO: 70 to 100% of the above SEQ ID NO: 70 to 100% of the specified fragment, domain, or variant thereof, or at least one of the specified fragments, domains, or variants thereof, in 40 to 99%. The percentage of such composition may be expressed by weight, volume, concentration, molar concentration, or molar concentration as a liquid or dry solution, mixture, suspension, emulsion, particles, powder, or colloid, as known in the art or as described herein.

[0117] Antibody composition containing further therapeutic active ingredients The antibody composition used in the method of the present invention may optionally further contain an effective amount of at least one compound or protein selected from at least one of the following: anti-infective agents, cardiovascular (CV) agonists, central nervous system (CNS) agents, autonomic nervous system (ANS) agents, respiratory agents, gastrointestinal (GI) agonists, hormonal agents, fluid or electrolyte balance agents, hematological agents, antitumor agents, immunomodulators, eye, ear or nose agents, topical agents, nutritional agents, etc. Such drugs, including their respective formulations, indications, dosages, and administrations as described herein, are well known in the art (see, for example, Nursing 2001 Handbook of Drugs, 21st edition, Springhouse Corp., Springhouse, PA, 2001; Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ; and Pharmcotherapy Handbook, Wells et al., Appleton & Lange, Stamford, CT, each incorporated herein by reference).

[0118] Examples of agents that can be combined with the antibody of the method of the present invention include, as an antiinfective agent, at least one selected from amoebic agents or at least one antiparasitic agent, anthelmintic agent, antifungal agent, antimalarial agent, antituberculosis agent or at least one anti-leprosy agent, aminoglycoside, penicillin, cephalosporin, tetracycline, sulfonamide, fluoroquinolone, antiviral agent, macrolide antiinfective agent, and various antiinfective agents. The hormonal agent may be at least one selected from corticosteroids, androgens or at least one anabolic steroid, estrogen or at least one progestin, gonadotropin, antidiabetic agent, or at least one glucagon, thyroid hormone, thyroid hormone antagonist, pituitary hormone, and parathyroid-like agent. At least one cephalosporin may be selected from cefaclor, cefadroxil, cefazolin sodium, cefdinir, cefepime hydrochloride, cefixime, cefmetazole sodium, cefonisid sodium, cefoperazone sodium, cefotaxime sodium, cefotetan disodium, cefoxitin sodium, cefpodoxime proxetil, cefprodil, ceftazidime, ceftibutene, ceftizoxime sodium, ceftriaxone sodium, cefuroxime axetil, cefuroxime sodium, cephalexin hydrochloride, cephalexin monohydrate, cefradin, and lorakarbef.

[0119] At least one corticosteroid may be selected from betamethasone, betamethasone acetate or sodium betamethasone phosphate, sodium betamethasone phosphate, cortisone acetate, dexamethasone, dexamethasone acetate, sodium dexamethasone phosphate, fludrocortisone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone cypionate, sodium hydrocortisone phosphate, sodium hydrocortisone succinate, methylprednisolone, methylprednisolone acetate, sodium methylprednisolone succinate, prednisolone, prednisolone acetate, sodium prednisolone phosphate, prednisolone tebutate, prednisone, triamcinolone, triamcinolone acetonide, and triamcinolone diacetate. At least one androgen or anabolic steroid may be selected from danazol, fluoxymesterone, methyltestosterone, nandrolone decanoate, nandrolone fenpropionate, testosterone, testosterone cypionate, testosterone enanthate, testosterone propionate, and the transdermal testosterone system.

[0120] At least one immunosuppressant may be selected from azathioprine, basiliximab, cyclosporine, daclizumab, lymphocyte immunoglobulin, muromonab-CD3, mycophenolate mofetil, mycophenolate mofetil hydrochloride, sirolimus, 6-mercaptopurine, methotrexate, mizoribine, and tacrolimus.

[0121] At least one topical antiinfective agent may be selected from acyclovir, amphotericin B, azelaic acid cream, bacitracin, butoconazole nitrate, clindamycin phosphate, clotrimazole, econazole nitrate, erythromycin, gentamicin sulfate, ketoconazole, mafenide acetate, metronidazole (topical), miconazole nitrate, mupirocin, naphthifine hydrochloride, neomycin sulfate, nitrofurazone, nystatin, silver sulfadiazine, terbinafine hydrochloride, terconazole, tetracycline hydrochloride, thioconazole, and tolnaftate. At least one scabies insecticide or lice killer may be selected from crotamiton, lindane, permethrin, and pyrethrin. At least one topical corticosteroid may be selected from betamethasone dipropionate, betamethasone valerate, clobetasol propionate, desonide, desoxymethasone, dexamethasone, dexamethasone sodium phosphate, diflorasone diacetate, fluocinolone acetonide, fluocinonide, flulandrenolide, fluticasone propionate, halcionide, hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone valerate, mometasone fluate, and triamcinolone acetonide. (See, for example, pages 1098-1136 of the Nursing 2001 Drug Handbook.)

[0122] An anti-IL-12 / IL-23p40 or IL-23 antibody composition comprises at least one anti-IL-12 / 23p40 or IL-23 antibody that is contacted with or administered to cells, tissues, organs, animals, or subjects requiring such regulation, treatment, or therapy, and optionally further comprises at least one TNF antagonist (e.g., but not limited to TNF chemical or protein antagonists, TNF monoclonal or polyclonal antibodies or fragments, soluble TNF receptors (e.g., p55, p70, or p85) or fragments, their fusion polypeptides, or small molecule TNF antagonists, e.g., TNF-binding protein I or II (TBP-1 or TBP-II), nerelimon mab) The composition or pharmaceutical composition may further include at least one of any suitable and effective amounts of any one selected from the following: mab), infliximab, etanacept, CDP-571, CDP-870, afelimomab, renercept, etc., antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, etanercept, sodium aurothiomalate, hydroxychloroquine sulfate, leflunomide, sulfasalazine), immunizing agents, immunoglobulins, immunosuppressants (e.g., azathioprine, basiliximab, cyclosporine, daclizumab), cytokines, or cytokine antagonists. Non-limiting examples of such cytokines include, but are not limited to, any of IL-1 to IL-23 (e.g., IL-1, IL-2, etc.). Suitable dosages are well known in the art. For example, see Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, CT (2000), and "PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000" Special Edition, Tarascon Publishing, Loma Linda, CA (2000), each of which is incorporated herein by reference in its entirety.

[0123] The anti-IL-12 / IL-23p40 or IL-23 antibody compound, composition, or mixture used in the method of the present invention may further include at least one of any suitable auxiliary agents, including but not limited to diluents, binders, stabilizers, buffers, salts, lipophilic solvents, preservatives, and adjuvants. A pharmaceutically acceptable auxiliary agent is preferred. Methods for preparing such sterile solutions and non-limiting examples thereof are well known in the art, for example, Gennaro, Ed., Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co. (Easton, PA), 1990. A pharmaceutically acceptable carrier suitable for the administration method, solubility, and / or stability of the anti-IL-12 / IL-23p40, fragment, or variant composition can be routinely selected, as is well known in the art or as described herein.

[0124] Useful pharmaceutical excipients and additives in this composition include, but are not limited to, proteins, peptides, amino acids, lipids, and carbohydrates (e.g., monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides, derivatized sugars such as alditol, aldonic acid, and esterified sugars, and polysaccharides or sugar polymers), which may be present alone or in combination, and may be present alone or in combination in amounts of 1 to 99.99% by weight or volume. Exemplary protein excipients include serum albumin such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, and casein. Representative amino acid / antibody components that can also function in buffering capacity include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, and aspartame. One preferred amino acid is glycine.

[0125] Suitable carbohydrate excipients for use in the present invention include, for example, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, and sorbose; disaccharides such as lactose, sucrose, trehalose, and cellobiose; polysaccharides such as raffinose, melegitose, maltodextrin, dextran, and starches; and algitols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), and myo-inositol. Preferred carbohydrate additives for use in the present invention are mannitol, trehalose, and raffinose.

[0126] Anti-IL-12 / IL-23p40 or IL-23 antibody compositions may also contain buffers or pH adjusters, typically the buffers being salts prepared from organic acids or bases. Typical buffers include organic acid salts such as citric acid, ascorbic acid, gluconic acid, carbonate, tartaric acid, succinic acid, acetic acid, or phthalic acid salts, Tris, tromethamine hydrochloride, or phosphate buffers. Organic acid salts such as citric acid are preferred buffers for use in this composition.

[0127] In addition, anti-IL-12 / IL-23p40 or IL-23 antibody compositions may contain polymer excipients / additives such as polyvinylpyrrolidone, Ficol (polymer sugar), dextrose (e.g., cyclodextrin such as 2-hydroxypropyl-β-cyclodextrin), polyethylene glycol, flavoring agents, antibacterial agents, sweeteners, antioxidants, antistatic agents, surfactants (e.g., polysorbates such as "TWEEN20" and "TWEEN80"), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0128] These and additional known pharmaceutically acceptable excipients and / or additives suitable for use in anti-IL-12 / IL-23p40 or IL-23 antibody, partial or variant compositions according to the present invention are known in the art and are listed, for example, in "Remington: The Science & Practice of Pharmacy," 19th ed., Williams & Williams, (1995) and "Physician's Desk Reference," 52nd ed., Medical Economics, Montvale, NJ (1998), the disclosures of which are incorporated herein by reference in their entirety. Preferred carrier or additive materials are carbohydrates (e.g., monosaccharides and alditols) and buffers (e.g., citric acid) or polymers. Exemplary carrier molecules are mucopolysaccharides and hyaluronic acid, which may be useful for intra-articular delivery.

[0129] formulation As described above, the present invention provides a stable formulation preferably comprising a phosphate buffer containing physiological saline or a selected salt, as well as a preservative solution and formulation containing a preservative, and a versatile preservative formulation suitable for pharmaceutical or veterinary use, comprising at least one anti-IL-12 / IL-23p40 or IL-23 antibody in a pharmaceutically acceptable formulation. The preservative formulation contains in an aqueous diluent a preservative optionally selected from the group consisting of at least one known preservative, i.e., at least one phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercury nitrate, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate), alkylparabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof. As is known in the art, 0.001 to 5%, or any range or value within that range, for example, 0.001, 0.003, 0.005, 0.009, 0.01, 0.02, 0.03, 0.05, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1 Any suitable concentration or mixture of 0.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.3, 4.5, 4.6, 4.7, 4.8, 4.9, etc., or any range or value within these, can be used.Non-limiting examples include: no preservatives added, 0.1-2% m-cresol (e.g., 0.2, 0.3, 0.4, 0.5, 0.9, 1.0%), 0.1-3% benzyl alcohol (e.g., 0.5, 0.9, 1.1, 1.5, 1.9, 2.0, 2.5%), 0.001-0.5% thimerosal (e.g., 0.005, 0.01%), 0.001-2.0% phenol (e.g., Examples include 0.05%, 0.25%, 0.28%, 0.5%, 0.9%, and 1.0%, and 0.0005-1.0% alkylparabens (multiple amounts possible) (for example, 0.00075%, 0.0009%, 0.001%, 0.002%, 0.005%, 0.0075%, 0.009%, 0.01%, 0.02, 0.05, 0.075, 0.09, 0.1%, 0.2%, 0.3%, 0.5%, 0.75%, 0.9%, and 1.0%).

[0130] As described above, the present invention uses an article comprising a packaging material and at least one vial containing a solution of at least one anti-IL-12 / IL-23p40 or IL-23 antibody having optionally formulated buffer and / or preservative in an aqueous diluent, wherein the packaging material includes a label indicating that such solution can be maintained for a period of 1, 2, 3, 4, 5, 6, 9, 12, 18, 20, 24, 30, 36, 40, 48, 54, 60, 66, 72 hours or more. The present invention further uses a product comprising a packaging material and a first vial containing lyophilized anti-IL-12 / IL-23p40 or IL-23 antibody and a second vial containing an aqueous diluent of the formulated buffer or preservative, wherein the packaging material includes a label instructing a subject to reconstitute the anti-IL-12 / IL-23p40 or IL-23 antibody with the aqueous diluent to form a solution that can be maintained for a period of 24 hours or more.

[0131] Anti-IL-12 / IL-23p40 or IL-23 antibodies used in accordance with the present invention may be produced by recombinant means such as from mammalian cells or transgenic formulations, as described herein or known in the art, or may be purified from other biological sources.

[0132] The anti-IL-12 / IL-23p40 or IL-23 antibody ranges from approximately 1.0 μg / mL to approximately 1000 mg / mL when reconstituted in a wet / dry system, but lower and higher concentrations are also possible, depending on the intended delivery vehicle, such as solution formulations, which may differ from transdermal patches, lung, transmucosal, or osmotic or micropump methods.

[0133] Preferably, the aqueous diluent further optionally comprises a pharmaceutically acceptable preservative. Preferred preservatives include those selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkylparabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof. The concentration of the preservative used in the formulation is sufficient to produce an antimicrobial effect. Such a concentration varies depending on the selected preservative and is readily determined by those skilled in the art.

[0134] Other excipients, such as isotonic agents, buffers, antioxidants, and preservative enhancers, can be optionally and preferably added to the diluent. Isotonic agents such as glycerin are commonly used at known concentrations. Preferably, physiologically tolerant buffers are added to provide improved pH control. Formulations can cover a wide pH range, such as about pH 4 to about pH 10, preferably about pH 5 to about pH 9, and most preferably about pH 6.0 to about pH 8.0. Preferably, the formulations of the present invention have a pH of about 6.8 to about 7.8. Suitable buffers include phosphate buffers, most preferably sodium phosphate, and especially phosphate-buffered saline (PBS).

[0135] Other additives, such as pharmaceutically acceptable solubilizers like Tween20 (polyoxyethylene(20) sorbitan monolaurate), Tween40 (polyoxyethylene(20) sorbitan monopalmitate), Tween80 (polyoxyethylene(20) sorbitan monooleate), Pluronic F68 (polyoxyethylene polyoxypropylene block copolymer), and PEG (polyethylene glycol), or nonionic surfactants such as polysorbate 20 or 80 or poloxamer 184 or 188, Pluronic® polyl, other block copolymers, and chelating agents such as EDTA and EGTA, can be optionally added to the formulation or composition to reduce aggregation. These additives are particularly useful when a pump or plastic container is used to dispense the formulation. The presence of pharmaceutically acceptable surfactants reduces the tendency of proteins to aggregate.

[0136] The formulation can be prepared by a process comprising mixing at least one anti-IL-12 / IL-23p40 or IL-23 antibody with a preservative selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkylparabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal or mixtures thereof) in an aqueous diluent. The mixing of at least one anti-IL-12 / IL-23p40 or IL-23 specific antibody and the preservative in the aqueous diluent is carried out using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a certain amount of at least one anti-IL-12 / IL-23p40 or IL-23 antibody in a buffer solution is combined with the desired preservative in a buffer solution sufficient to provide the desired concentration of protein and preservative. Variations of this process will be recognized by those skilled in the art. For example, the order in which the constituent components are added, whether or not additional additives are used, and the temperature and pH during formulation preparation are all factors that can be optimized in relation to the dosage concentration and means of administration used.

[0137] The formulation can be provided to a target as a clear solution, or as a dual vial containing a vial of lyophilized anti-IL-12 / IL-23p40 or IL-23 specific antibody, which is reconstituted in a second vial containing water, a preservative and / or excipient, preferably a phosphate buffer and / or physiological saline, and a selected salt in an aqueous diluent. Both the single solution vial and the dual vial requiring reconstitution can be reused multiple times and may be sufficient for one or more treatment cycles for a target, thus providing a more convenient dosing regimen than currently available.

[0138] This product is useful for administration over a period ranging from immediate to 24 hours or more. Therefore, the product claimed by this invention offers significant benefits to the target audience. The formulation of this invention can be safely stored as desired at temperatures of approximately 2°C to approximately 40°C and can maintain the biological activity of the protein for extended periods; therefore, the packaging label can indicate that the solution can be retained and / or used for 6, 12, 18, 24, 36, 48, 72, or 96 hours or more. When using a stored diluent, such label may include use for up to 1 to 12 months, 6 months, 1.5 years, and / or 2 years.

[0139] Solutions of anti-IL-12 / IL-23p40 or IL-23 specific antibodies can be prepared by a process involving mixing at least one antibody in an aqueous diluent. The mixing is carried out using conventional dissolution and mixing procedures. To prepare a suitable diluent, for example, a certain amount of at least one antibody in water or a buffer is combined with a protein of the desired concentration and, optionally, an amount sufficient to provide a preservative or buffer. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of components, the use of additional additives, and the temperature and pH during formulation preparation are all factors that can be optimized with respect to the dosage concentration and means of administration used.

[0140] The claimed product may be provided to a target as a clear solution or as a dual vial containing at least one lyophilized vial of an anti-IL-12 / IL-23p40 or IL-23 specific antibody, which is reconstituted in a second vial containing an aqueous diluent. Both the single-solution vial and the dual vial requiring reconstitution can be reused multiple times and may be sufficient for one or more treatment cycles for a target, thus providing a more convenient dosing regimen than currently available.

[0141] The claimed product can be indirectly provided to the target by providing a dual vial containing a clear solution or an aqueous diluent, which is reconstituted in a second vial containing a lyophilized vial of at least one anti-IL-12 / IL-23p40 or IL-23 specific antibody, to a pharmacy, clinic, or other such institution and facility. The clear solution in this case may be up to 1 liter or even more, and smaller amounts of the at least one antibody solution can be taken from this larger container one or more times and transferred to smaller vials, which can then be provided to customers and / or the target by the pharmacy or clinic.

[0142] Approved devices including single vial systems include pen-type injector devices for delivering solutions, such as BD Pens, BD Autojector®, Humaject®, NovoPen®, B-DPen®, AutoPen®, and OptiPen®, GenotropinPen®, Genotronorm Pen®, Humatro Pen®, Reco-Pen®, Roferon Pen®, Biojector®, Iject®, J-tip Needle-Free Injector®, Intraject®, Medi-Ject®, Smartject®, etc. (Becton Dickensen (Franklin) Suitable devices include those manufactured or developed by Lakes, NJ (www.bectondickenson.com), Disetronic (Burgdorf, Switzerland (www.disetronic.com)), Bioject (Portland, Oregon (www.bioject.com)), National Medical Products, Weston Medical (Peterborough, UK (www.weston-medical.com)), and Medi-Ject Corp (Minneapolis, MN (www.mediject.com)), and similar devices. Approved devices, including dual vial systems, include pen-type syringe systems for reconstituting lyophilized drugs in a cartridge for delivering reconstituted solutions, such as HumatroPen®. Examples of other suitable devices include pre-filled syringes, auto-injectors, needleless syringes, and needleless IV infusion sets.

[0143] The product may include packaging materials. The packaging materials provide information required by regulatory authorities, as well as conditions for the use of the product. The packaging materials of the present invention, relating to two vial-type wet / dry products, provide instructions, where applicable, to reconstitute at least one anti-IL-12 / IL-23p40 or IL-23 antibody with an aqueous diluent to form a solution, and to use this solution over a period of 2 to 24 hours or more. For single-vial solution products, pre-filled syringes, or auto-injectors, the label indicates that such solution can be used over a period of 2 to 24 hours or more. The product is useful for human pharmaceutical product applications.

[0144] The formulations used in the method of the present invention can be prepared by a process comprising mixing anti-IL-12 / IL-23p40 with a selected buffer, preferably physiological saline or a phosphate buffer containing a selected salt. The mixing of the anti-IL-12 / IL-23p40 antibody and the buffer in an aqueous diluent is carried out using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a certain amount of at least one antibody in water or buffer is combined with the desired buffer in a sufficient amount of water to provide the desired concentration of protein and buffer. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of components, the use or non-use of additional additives, and the temperature and pH during formulation preparation are all factors that can be optimized with respect to the dosage concentration and means of administration used.

[0145] The present invention provides a method for pharmaceutical compositions comprising various formulations useful and acceptable for administration to human or animal subjects. Such pharmaceutical compositions are prepared using "standard condition" water as a diluent and routine methods well known to those skilled in the art. For example, buffering components such as histidine and histidine monohydrochloride hydrate are first provided, followed by the addition of a suitable non-final volume of aqueous diluent, sucrose, and polysorbate 80 under "standard condition." Isolated antibodies can then be added. Finally, the volume of the pharmaceutical composition is adjusted to the desired final volume under "standard condition" conditions using water as a diluent. Those skilled in the art will recognize several other methods suitable for preparing pharmaceutical compositions.

[0146] A pharmaceutical composition may contain each component by a specified mass per unit volume of water, or may be an aqueous solution or suspension having a pH specified at “standard conditions.” As used herein, “standard conditions” means a temperature of 25°C ± 2°C and a pressure of 1 atm. The term “standard conditions” is not used in the art to refer to a single set of temperatures or pressures recognized by the art, but rather to specify a reference state of temperature and pressure used to describe a solution or suspension containing a particular composition under reference “standard conditions” conditions. This is because the volume of a solution is, in part, a function of both temperature and pressure. Those skilled in the art will recognize that pharmaceutical compositions equivalent to those disclosed herein may be produced at other temperatures and pressures. Whether such pharmaceutical compositions are equivalent to those disclosed herein should be determined under the “standard conditions” conditions defined above (e.g., 25°C ± 2°C and a pressure of 1 atm).

[0147] Importantly, such a pharmaceutical composition may contain "approximately" a certain mass of a component (e.g., "approximately 0.53 mg of L-histidine") or have an approximately certain pH value per unit volume of the pharmaceutical composition. The mass of a component or the pH value present in the pharmaceutical composition is "approximately" a given value such that the isolated antibody is present in the pharmaceutical composition, or that the isolated antibody present in the pharmaceutical composition can bind to the peptide chain after it has been removed from the pharmaceutical composition (e.g., by dilution). In other words, the value such as the mass of a component or the pH value is "approximately" a given value such that the binding activity of the isolated antibody is maintained and detectable after it has been added to the pharmaceutical composition.

[0148] Competitive binding analysis is performed to determine whether IL-12 / IL-23p40 or IL-23-specific mAbs bind to similar or different epitopes and / or compete with each other. The mAbs are individually coated onto an ELISA plate. Competing mAbs are added, followed by biotinylated hrIL-12 or IL-23. For a positive control, the same mAb used in the coating may be used as a competing mAb ("self-competition"). IL-12 / IL-23p40 or IL-23 binding is detected using streptavidin. These results indicate whether the mAb recognizes similar or partially overlapping epitopes on IL-12 / IL-23p40 or IL-23.

[0149] In one embodiment of the pharmaceutical composition, the isolated antibody concentration is approximately 77 to 104 mg per 1 mL of the pharmaceutical composition. In another embodiment of the pharmaceutical composition, the pH is approximately 5.5 to 6.5.

[0150] The stable or preserved formulation can be provided to the patient as a clear solution, or as a dual vial containing at least one lyophilized vial of anti-IL-12 / IL-23p40, which is reconstituted in a second vial containing a preservative or buffer and excipients in an aqueous diluent. Both the single solution vial and the dual vial requiring reconstitution can be reused multiple times and may be sufficient for one or more target treatment cycles, thus providing a more convenient dosing regimen than currently available.

[0151] Other formulations or methods for stabilizing anti-IL-12 / IL-23p40 antibodies may be other than clear solutions of lyophilized powder containing the antibody. Opaque solutions include formulations comprising a suspension of microparticles, the microparticles being compositions containing the anti-IL-12 / IL-23p40 antibody within structures of various dimensions, variously known as microspheres, fine particles, nanoparticles, nanospheres, or liposomes. Such relatively homogeneous, essentially spherical microparticle formulations containing an active agent can be formed by contacting an aqueous phase containing an active agent and polymer with a non-aqueous phase, as taught in U.S. Patent No. 4,589,330, and then evaporating the non-aqueous phase to induce accretion of particles from the aqueous phase. Porous microparticles can be prepared by using a first phase containing an active agent and polymer dispersed in a continuous solvent, as taught in U.S. Patent No. 4,818,542, and removing the solvent from the suspension by lyophilization or dilution-extraction-precipitation. Polymers preferred for these preparations include gelatin agar, starch, arabinogalactan, albumin, collagen, polyglycolic acid, polylactic acid, glycoside-L(-)lactide, poly(epsilon-caprolactone, poly(epsilon-caprolactone-CO-lactic acid), poly(epsilon-caprolactone-CO-glycolic acid), poly(β-hydroxybutyric acid), polyethylene oxide, polyethylene, poly(alkyl-2-cyanoacrylate), poly(hydroxyethyl methacrylate), polyamide, poly(amino acid), poly(2-hydroxyethyl methacrylate) The polymer is a natural or synthetic copolymer or polymer selected from the group consisting of roxyethyl DL-aspartamide, poly(ester urea), poly(L-phenylalanine / ethylene glycol / 1,6-diisocyanatohexane), and poly(methyl methacrylate). Particularly preferred polymers are polyesters such as polyglycolic acid, polylactic acid, glycolide-L(-)lactide poly(epsilon-caprolactone), poly(epsilon-caprolactone-CO-lactic acid), and poly(epsilon-caprolactone-CO-glycolic acid).Suitable solvents for dissolving polymers and / or active substances include water, hexafluoroisopropanol, methylene chloride, tetrahydrofuran, hexane, benzene, or hexafluoroacetone sesquihydrate. The process of dispersing the active substance-containing phase into the second phase may include a step of forcing the first phase through an orifice in a nozzle under pressure to induce droplet formation.

[0152] Dry powder formulations can be obtained as a result of processes other than lyophilization, such as spray drying, solvent extraction by evaporation, or precipitation of a crystalline composition followed by one or more steps to remove an aqueous or non-aqueous solvent. The preparation of spray-dried antibody formulations is taught in U.S. Patent No. 6,019,968. Antibody-based dry powder compositions can be produced by spray-drying a solution or slurry of antibody in a solvent, and optionally excipients, under conditions for providing a respiration-drying powder. Solvents may include polar compounds that dry easily, such as water and ethanol. Antibody stability can be enhanced by performing the spray-drying procedure in the absence of oxygen, for example, under a nitrogen blanket, or by using nitrogen as the drying gas. Another relatively dry formulation is a dispersion of multiple porous microstructures dispersed in a suspension medium, typically containing a hydrofluoroalkane propellant, as taught in International Publication No. 9916419. The stabilized dispersion can be administered to the target lung using a metered-dose inhaler. Equipment useful in the commercial manufacture of spray-dried drugs is manufactured by Buchi Ltd. or Niro Corp.

[0153] The anti-IL-12 / IL-23p40 in any of the stable or preserved formulations or solutions described herein can be administered to a subject in accordance with the present invention by various delivery methods, such as SC or IM injection, transdermal, transpulmonary, transmucosal, implantation, osmotic pump, cartridge, micropump, or other means as understood by those skilled in the art, as is well known in the art.

[0154] Applications in treatment The present invention also provides a method for modulating or treating ulcerative colitis in cells, tissues, organs, animals, or subjects by administering or contacting, for example, cells, tissues, organs, animals, or subjects with at least one IL-23 antibody of the present invention, as is known in the art or as described herein, with a therapeutically effective amount of IL-12 / IL-23p40 or IL-23 specific antibody.

[0155] Any method of the present invention may include administering an effective amount of a composition or pharmaceutical composition containing IL-12 / IL-23p40 to cells, tissues, organs, animals, or subjects requiring such regulation, treatment, or therapy. Such methods may further include, optionally, concurrent or combination therapy for the treatment of such diseases or disorders, wherein the administration of at least one IL-12 / IL-23p40, a specific portion thereof, or a variant thereof, is equivalent to administering at least one TNF antagonist (e.g., chemical or protein-based TNF antagonists, TNF monoclonal or polyclonal antibodies or fragments, soluble TNF receptors (e.g., p55, p70, or p85) or fragments, their fusion polypeptides, or small molecule TNF antagonists, e.g., , TNF-binding protein I or II (TBP-1 or TBP-II), nerimomab, infliximab, etanercept (Enbrel®), adalimumab (Humira®), CDP-571, CDP-870, afelimomab, renercept, etc., antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, sodium aurothiomalate, hydroxychloroquine sulfate, leflunomide, sulfasalazine), muscle relaxants, narcotics, non-steroidal anti-inflammatory drugs (NSAIDs)Drugs, NSAIDs (e.g., 5-aminosalicylates), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blocking agents, antibacterial agents (e.g., aminoglycosides, antifungals, antiparasitic agents, antivirals, carbapenams, cephalosporins, fluoroquinolones, macrolides, penicillin, sulfonamides, tetracyclines, and other antibacterial agents), psoriasis medications, corticosteroids, anabolic steroids, diabetes-related medications, minerals, nutritional supplements, thyroid medications, vitamins, calcium-related hormones, antidiarrheals, antitussives, antiemetics, antitumor drugs, laxatives, anticoagulants, erythropoietin (e.g., epoetin alfa), filgrastim (e.g., G-CSF, Neupogen), salgramostim (GM-CSF, L The treatment further comprises administering, simultaneously with, and / or after, at least one selected from eukine, immunoconjugates, immunoglobulins, immunosuppressants (e.g., basiliximab, cyclosporine, daclizumab), growth hormone, hormone replacement therapy, estrogen receptor modulators, mydriatics, ciliary muscle paralyzers, alkylating agents, antimetabolites, mitotics, radiopharmaceuticals, antidepressants, antimanic drugs, antipsychotics, anxiolytics, hypnotics, sympathomimetic agents, stimulants, donepezil, tacrine, asthma treatments, beta-agonists, inhaled corticosteroids, leukotriene inhibitors, methylxanthines, cromolyn, epinephrine or analogs, dorunase alfa (Pulmozyme), cytokines or cytokine antagonists. Preferred dosages are well known in the art. For example, Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, CT (2000), PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, CA (2000), Nursing 2001 Handbook of Drugs, 21st edition, Springhouse. Corp., Springhouse, PA, 2001, Health Professional's Drug GuideSee 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ, each of these references is incorporated herein by reference in whole.

[0156] Treatment The treatment of ulcerative colitis is influenced by administering an effective dose or amount of an anti-IL-12 / 23p40 composition to the patient in need. The dose administered may vary depending on known factors such as the pharmacodynamic characteristics of the particular drug, its method and route of administration, the recipient's age, health status and weight, the nature and severity of symptoms, the type and frequency of concurrent treatments, and the desired effect. In some cases, repeated dosing, i.e., repeated individual doses of a specific monitored or measured dose, may be necessary to achieve the desired therapeutic dose, in which case the individual doses are repeated until the desired daily dose or effect is obtained.

[0157] In one exemplary regimen providing safe and effective treatment for severe, active UC in patients in need, a total dose of approximately 130 mg of anti-IL-12 / IL-23p40 antibody is administered intravenously to the patient with each dose. For example, the total volume of the administered composition is appropriately adjusted to provide the target dose of antibody to the patient, such as 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, or 180 mg per dose.

[0158] In one exemplary regimen providing safe and effective treatment for severe, active UC in patients in need, a total dose of approximately 6.0 ± 1.5 mg / kg of anti-IL-12 / IL-23p40 antibody is administered intravenously to the patient with each dose. For example, the total amount of the composition administered per kg of body weight of the patient per dose is appropriately adjusted to provide the target dose of antibody to the patient, such as 3.0 mg / kg, 3.5 mg / kg, 4.0 mg / kg, 4.5 mg / kg, 5.0 mg / kg, 5.5 mg / kg, 6.0 mg / kg, 6.5 mg / kg, 7.0 mg / kg, 7.5 mg / kg, 8.0 mg / kg, 8.5 mg / kg, or 9.0 mg / kg.

[0159] The total dose of anti-IL-12 / IL-23p40 antibody administered to the target in a single dose can be given by intravenous infusion over a period of approximately 30 to 180 minutes, preferably 60 to 120 minutes (for example, 30, 60, 90, 120, 150, or 180 minutes).

[0160] In another exemplary regimen providing safe and effective treatment for severe, active UC in patients in need, a total dose of approximately 90 mg of anti-IL-12 / IL-23p40 antibody is administered subcutaneously to the patient with each dose. For example, the total amount of the composition administered is appropriately adjusted to provide the target dose of antibody to the patient, such as 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, or 140 mg per dose. The target dose per dose can be administered as a single subcutaneous injection or as multiple subcutaneous injections, for example, one, two, three, four, five, or more subcutaneous injections.

[0161] The total dose of anti-IL-12 / IL-23p40 antibody can be administered over periods of one day, one week, one month, six months, one year, two years, or longer, such as once daily, once weekly, once monthly, or once every six months. Multiple doses of anti-IL-12 / IL-23p40 antibody at the total doses described herein can be administered to subjects requiring it.

[0162] Dosage forms (compositions) suitable for internal administration generally contain approximately 0.001 milligrams to approximately 500 milligrams of the active ingredient per unit or container.

[0163] For parenteral administration, antibodies may be formulated as solutions, suspensions, emulsions, particles, powders, or lyophilized powders, either in combination with or separately provided with a pharmaceutically acceptable parenteral vehicle. Examples of such vehicles include water, physiological saline, Ringer's solution, dextrose solution, and 1-10% human serum albumin. Non-aqueous vehicles such as liposomes and non-volatile oils may also be used. The vehicle or lyophilized powder may contain additives to maintain isotonicity and chemical stability (e.g., sodium chloride and mannitol for isotonicity, and buffers and preservatives for chemical stability). The formulations are sterilized by known or preferred techniques.

[0164] Suitable pharmaceutical carriers are described in the latest edition of Remington's Pharmaceutical Sciences, by A. Osol, which is the standard reference text in this field.

[0165] Many known and developed methods can be used in accordance with the present invention to administer a pharmaceutically effective amount of IL-12 / IL-23p40 antibody. The IL-12 / IL-23p40 or IL-23 antibody of the present invention can be delivered in a carrier as a solution, emulsion, colloid, or suspension, or as a dry powder, by inhalation or by any of the various devices and methods suitable for administration by methods described herein or known in the art.

[0166] Parenteral formulations may contain sterile water or saline solution, polyalkylene glycols such as polyethylene glycol, vegetable oils, hydrogenated naphthalene, etc., as common excipients. Aqueous or oily suspensions for injection can be prepared by using appropriate emulsifiers or humectants and suspensions according to known methods. Injectable formulations may be non-toxic parenterally administered diluents such as aqueous solutions, sterile injection solutions, or suspensions in solvents. Suitable vehicles or solvents include water, Ringer's solution, and isotonic saline solution, and sterile non-volatile oils can be used as ordinary solvents or suspension solvents. For these purposes, all kinds of non-volatile oils and fatty acids, including natural, synthetic, or semi-synthetic fatty oils or fatty acids, and natural, synthetic, or semi-synthetic monoglycerides, diglycerides, or triglycerides, can be used. Parenteral administration is known in the art and includes, but is not limited to, conventional injection methods, gas-pressurized needleless injection devices such as those described in U.S. Patent No. 5,851,198, and laser drilling devices such as those described in U.S. Patent No. 5,839,446, all of which are incorporated herein by reference.

[0167] alternative delivery The present invention further relates to the administration of anti-IL-12 / IL-23p40 or IL-23 antibodies by parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intra-bronchial, intra-abdominal, intracapsular, intra-cartilaginous, intra-sinusinal, intra-cavitary, intra-cerebellar, intra-cervical, intra-colon, intra-cervical, intra-stomach, intra-hepatic, intra-myocardium, intra-osseous, intra-pelvic, intra-pericardial, intra-abdominal, intra-pleural, intra-prostate, intra-lung, intra-rectal, intra-renal, intra-retinal, intra-spinal cord, intra-synovial sac, intra-thoracic, intra-uterine, intra-bladder, intra-focal, bolus, intra-vaginal, rectal, intra-oral cavity, sublingual, intra-nasal, or percutaneous means. Anti-IL-12 / IL-23p40 or IL-23 antibody compositions may be used parenterally (subcutaneous, intramuscular, or intravenously) or in any other form, particularly in the form of liquid solutions or suspensions, particularly in semi-solid forms such as creams and suppositories, for vaginal or rectal administration, in the form of tablets or capsules, but not limited to these, for oral or sublingual administration, or in the form of powders, nasal drops, aerosols, or certain drugs, but not limited to these, for intranasal use, or with chemical enhancers such as dimethyl sulfoxide to either modify skin structure or increase drug concentration in transdermal patches (Junginger et al., In "Drug Permeation Enhancement"; Hsieh, DS, Eds., pp. 59-90 (Marcel Dekker, Inc. New York) These can be prepared for transdermal use in the form of gels, ointments, lotions, suspensions, or patch delivery systems, etc., using an oxidizing agent that enables the application of formulations containing proteins and peptides to the skin (International Publication No. 98 / 53847), or the application of an electric field to create a transient transport pathway such as electroporation, or to increase the mobility of charged drugs through the skin such as iontophoresis, or the application of ultrasound such as sonication (U.S. Patents No. 4,309,989 and 4,767,402), but not limited to these applications (the above publications and patents are incorporated herein by reference in their entirety).

[0168] Embodiment The present invention also provides the following non-limiting embodiments. 1. A method for treating moderate to severe active ulcerative colitis (UC) in a subject in need, comprising administering a pharmaceutical composition to the subject containing a clinically proven safe and clinically proven effective amount of anti-IL-12 / IL-23p40 antibody, wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the complementarity-determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, the CDRH2 amino acid sequence of SEQ ID NO: 2, and the CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region The antibody comprises the complementarity-determining region light chain 1 (CDRL1) amino acid sequence of sequence number 4, the CDRL2 amino acid sequence of sequence number 5, and the CDRL3 amino acid sequence of sequence number 6. The antibody is administered intravenously to the subject, preferably at week 0 of treatment, at a dose of approximately 6.0 mg / kg of the subject's body weight or 130 mg per dose, and subcutaneously to the subject, preferably at week 8 of treatment, at a dose of 90 mg per dose. The antibody is then administered at a maintenance dose every 8 weeks after treatment at week 8, or every 12 weeks after treatment at week 8, and the subject... (a) Symptomatic remission, (b) Partial Mayo remission, (c) Mayo rectal bleeding subscore is 0, (d) Mayo bowel movement frequency subscore is 0 or 1, (e) The average absolute number of feces decreases by at least 3, (f) Reduction in the use and / or dosage of corticosteroids, (g) Corticosteroid-free symptomatic remission, (h) Corticosteroid-free partial Mayo remission, (i) Normalization of fecal lactoferrin, (j) Normalization of fecal calprotectin levels, (k) Total score on the Inflammatory Bowel Disease Questionnaire (IBDQ) improves by 16 points or more from baseline during the induction period. (l) IBDQ remission, (m)SF-36 PCS score improved by 5 points or more from the baseline during the introduction period, (n) SF-36 MCS score improves by 5 points or more from baseline during the introduction phase. A method for determining whether a responder to treatment at week 92 is selected from a group consisting of the following criteria, based on meeting one or more clinical endpoints. 2. The method according to Embodiment 1, wherein the antibody comprises the heavy chain variable region of the amino acid sequence of SEQ ID NO: 7 and the light chain variable region of the amino acid sequence of SEQ ID NO: 8. 3. The method according to Embodiment 1, wherein the antibody comprises a heavy chain of the amino acid sequence of SEQ ID NO: 10 and a light chain of the amino acid sequence of SEQ ID NO: 11. 4. The method according to any one of Embodiments 1 to 3, wherein the subject has previously failed or had poor tolerance to at least one therapy selected from the group consisting of anti-TNF, vedolizumab, corticosteroids, azathioprine (AZA), and 6-mercaptopurine (6MP), or the subject has shown corticosteroid dependence. 5. The method according to any one of Embodiments 1 to 3, wherein the subject is in corticosteroid-free clinical remission for at least 92 weeks from week 0. 6. The method according to any one of Embodiments 1 to 3, wherein the subject is identified as a responder to antibody treatment and has endoscopic healing that lasts for at least 92 weeks from week 0. 7. The method according to any one of Embodiments 1 to 6, wherein the pharmaceutical composition for intravenous administration further comprises a solution containing 10 mM L-histidine, 8.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80, 0.4 mg / mL L-methionine, and 20 μg / mL EDTA disodium salt dihydrate at pH 6.0. 8. The method according to any one of Embodiments 1 to 7, wherein the pharmaceutical composition for subcutaneous administration further comprises a solution containing 6.7 mM L-histidine, 7.6% (w / v) sucrose, and 0.004% (w / v) polysorbate 80 at pH 6.0.

[0169] While the present invention has been described in general terms, the same will be more readily apparent by referring to the following examples, which are provided as examples but are not intended to limit the scope of the invention. Furthermore, details of the present invention are illustrated by the following non-limiting examples. All disclosures by reference herein are expressly incorporated herein by reference. [Examples]

[0170] Example 1: Induction study of ustekinumab in the treatment of ulcerative colitis in humans The following multicenter, randomized, double-blind, placebo-controlled clinical trial was conducted in adult men and women with moderate to severe active ulcerative colitis (UC): A phase 3, randomized, double-blind, placebo-controlled, parallel-group, multicenter trial to evaluate the safety and efficacy of ustekinumab induction and maintenance therapy in subjects with moderate to severe active ulcerative colitis.

[0171] Overall theoretical basis A study was conducted to evaluate the efficacy of intravenous (IV) ustekinumab in patients with moderate to severe active ulcerative colitis who had shown insufficient response to or poor tolerance to conventional therapy (corticosteroids or 6-mercaptopurine / azathioprine [6-MP / AZA]) or biological therapy (TNF antagonists and / or integrin antagonists, vedolizumab). Patients received a single IV dose of 130 mg, a single IV dose of 6 mg / kg, or placebo at week 0. Patients who had not shown a clinical response at week 8 received an additional IV or subcutaneous (SC) dose at week 8.

[0172] the purpose The primary objectives of this study included (1) evaluating the efficacy of ustekinumab in inducing clinical remission in patients with moderate to severe active UC, and (2) evaluating the safety of IV ustekinumab in patients with moderate to severe active UC.

[0173] Secondary objectives of this study included: (1) evaluating the efficacy of IV ustekinumab in inducing endoscopic healing (i.e., improvement in endoscopic observation of the mucosa) in subjects with moderate to severe active UC; (2) evaluating the efficacy of IV ustekinumab in inducing clinical response in subjects with moderate to severe active UC; (3) evaluating the impact of IV ustekinumab on disease-specific health-related quality of life; (4) evaluating the efficacy of ustekinumab treatment for mucosal healing (i.e., endoscopic and histological healing); (5) evaluating the efficacy of induction therapy with IV ustekinumab by the status of biological failure; and (6) evaluating the pharmacokinetics (PK), immunogenicity, and pharmacodynamics (PD) of ustekinumab induction therapy in subjects with moderate to severe active UC, including changes in C-reactive protein (CRP), fecal calprotectin, fecal lactoferrin, and other PD biomarkers.

[0174] The exploratory objectives of the study included (1) evaluating responses using the Mayo score without the Comprehensive Physician Assessment (PGA) subscore, and (2) evaluating the performance of the Bristol Stool Form Scale (BSFS) score.

[0175] Experimental Design The Phase 3 development program for ustekinumab included two separate trials: an induction trial and a maintenance trial. In the induction trial, participants were randomized at week 0 to one of three treatment groups: placebo, low-dose ustekinumab, and high-dose ustekinumab. At week 8, all participants were evaluated for the primary endpoint of clinical remission and clinical response. Participants who achieved a clinical response at week 8 were eligible to enter the maintenance trial. Participants who did not achieve a clinical response at week 8 received a second dose of ustekinumab at week 8 of treatment.

[0176] At week 16, subjects who had not achieved a clinical response at week 8 were re-evaluated for their clinical response. Subjects who achieved a clinical response at week 16 were eligible to enter the maintenance trial. Subjects who did not achieve a clinical response at week 16 were not eligible to enter the maintenance trial and visited the hospital for safety follow-up approximately 20 weeks after the last dose of the study drug (week 8).

[0177] Patients who demonstrated a clinical response to IV ustekinumab during induction constituted the primary population for the maintenance trial. The maintenance trial is a randomized treatment discontinuation trial designed to evaluate maintenance therapy using SC ustekinumab and is currently ongoing.

[0178] Dosage and administration Participants received either a single IV dose of ustekinumab or placebo during week 0 of the study. The induction antibody levels based on the administered dose are as follows: ● Ustekinumab, low fixed dose of 130 mg ● Ustekinumab, high dose based on a body weight range of approximately 6 mg / kg: ○ Ustekinumab 260 mg (body weight ≤ 55 kg) ○ Ustekinumab 390mg (body weight > 55kg, but ≤ 85kg) ○ Ustekinumab 520 mg (body weight > 85 kg).

[0179] In subjects who did not show a clinical response, a second dose of ustekinumab was administered at week 8. The test antibodies with the second dose administered were as follows: ●Individuals randomized to placebo at week 0 received a single dose of ustekinumab approximately 6 mg / kg IV plus placebo SC (to maintain blindness) at week 8. ●Individuals randomized to ustekinumab at week 0 received a single dose of ustekinumab 90 mg SC + placebo IV (to maintain blindness) at week 8.

[0180] Safety evaluation Safety was evaluated based on adverse events (AEs) and laboratory test results (i.e., hematology and serology). Adverse events were either spontaneously reported by subjects or obtained through non-inducing interviews with subjects at the time of trial visit. Safety evaluation included the following laboratory tests. ●Hematology: Hemoglobin (Hb), hematocrit, red blood cell count, white blood cell (WBC) count, and platelets. ● Serum chemistry: Sodium, potassium, chloride, blood urea nitrogen (BUN), creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin and direct bilirubin, alkaline phosphatase, calcium, phosphate, albumin, total protein. ●Screening: Serological tests for human immunodeficiency virus antibodies, hepatitis C virus (HCV) antibodies, hepatitis B virus (HBV) antibodies, hepatitis B surface antigen, HBV surface antibody (anti-HB), and total HBV core (anti-HB) antibody tests, QuantiFERON-TB Gold test, pregnancy test (beta-human chorionic gonadotropin [βHCG]).

[0181] Pharmacokinetics Blood samples for measuring serum ustekinumab concentrations were collected at week 0 (before and after infusion) and at weeks 2, 4, and 8. Serum ustekinumab concentrations were analyzed using the validated electrochemiluminescence immunoassay (ECLIA) method on the Mesoscale Discovery (MSD®) platform (Gaithersburg, MD, USA). The lowest quantifiable concentration in the samples obtained by the ECLIA method using the MSD platform was 0.1688 μg / mL.

[0182] immunogenicity Antibodies against ustekinumab were evaluated using serum samples collected from all subjects. Analysis of antibodies against ustekinumab was performed using a confirmed, efficacy-validated drug resistance, electrochemiluminescence immunoassay (ECLIA), which captures and detects the immune response induced by ustekinumab. Antibody titers were determined for all subjects with antibodies against ustekinumab, and the neutralizing antibody (Nab) status of anti-drug antibody-positive samples was determined.

[0183] Evaluation of effectiveness Efficacy assessments were collected throughout the entire trial. The Mayo score and partial Mayo score, the Ulcerative Colitis Endoscopic Index of Severity (UCEIS), the Bristol Stool Form Scale (BSFS) score, C-reactive protein (CRP), fecal lactoferrin, fecal calprotectin, the Inflammatory Bowel Disease Questionnaire (IBDQ), the 36-item Short Form Health Questionnaire (SF-36), and the EuroQoL-5D Health Questionnaire were all evaluated to determine efficacy. Efficacy criteria were defined as follows: ●Clinical remission (world application): Mayo score of 2 or less, with no individual subscores exceeding 1. ●Clinical remission (US application): Absolute fecal count of 3 or more, rectal bleeding subscore of 0, and Mayo endoscopy subscore of 0 or 1. ●Clinical response: A decrease of 30% or more from baseline in the Mayo score and 3 points or more, or a decrease of 1 or more from baseline in the rectal bleeding subscore, or a rectal bleeding subscore of 0 or 1. ● Endoscopic healing (i.e., improvement in endoscopic observation of the mucosa): Mayo endoscopic subscore of 0 or 1. ● Histological healing: Defined based on the Geboes score, with epithelial neutrophils greater than 0 and less than 5%, and without crypt destruction, erosion, ulceration, or granulation tissue. ● Mucosal healing: Both endoscopic and histological healing. ● Normal or inactive mucosal disease: Mayo endoscopic subscore is 0. ●Symptomatic remission: Mayo bowel movement frequency subscore is 0 or 1, and rectal bleeding subscore is 0. ●Normalization of CRP concentration: CRP concentration is 3 mg / L or less. ●Normalization of fecal lactoferrin concentration: Fecal lactoferrin concentration is 7.24 μg / g or less. ● Normalization of fecal calprotectin concentration: Fecal calprotectin concentration is 250 mg / kg or less. ● Modified Mayo score response: ○Definition 1: A decrease of 2 points or more and 35% or more in the modified Mayo score, and a decrease of 1 or more in the rectal bleeding subscore, or a rectal bleeding subscore of 0 or 1. ○Definition 2: A decrease of 2 points or more and 30% or more in the modified Mayo score, and a decrease of 1 or more in rectal bleeding, or a rectal bleeding score of 0 or 1.

[0184] Safety results Both intravenous ustekinumab doses of approximately 6 mg / kg and 130 mg demonstrated generally favorable tolerability and a safety profile largely comparable to placebo throughout the 8-week period. In the safety analysis set of 960 patients, one or more treatment-related adverse events (AEs) were reported in 50.0%, 41.4%, and 48.0% of patients in the approximately 6 mg / kg, 130 mg, and placebo groups, respectively, throughout the 8-week period. Serious adverse events (SAEs) were reported in 3.1%, 3.7%, and 6.6% of patients in the approximately 6 mg / kg, 130 mg, and placebo groups, respectively, throughout the 8-week period.

[0185] The incidence of adverse events (AEs) within one hour of infusion was approximately 0.9% in the 6 mg / kg, 2.2% in the 130 mg, and 1.9% in the placebo group, respectively.

[0186] The proportion of subjects with one or more infections was 15.3% in the 6 mg / kg, 15.9% in the 130 mg, and 15.0% in the placebo group, respectively. Serious infections were reported in 0.3% in the 6 mg / kg, 0.6% in the 130 mg, and 1.3% in the placebo group, respectively.

[0187] Pharmacokinetic results Serum samples were collected at week 0 (pre-administration), week 0 (1 hour after administration), week 2, week 4, and week 8. For subjects randomized to ustekinumab treatment, a single IV infusion of ustekinumab was administered as an adjusted dose based on body weight of approximately 6 mg / kg (i.e., 260 mg for subjects with a body weight of ≤55 kg, 390 mg for subjects with a body weight of >55 kg and ≤85 kg, or 520 mg for subjects with a body weight of >85 kg), or as a fixed dose of 130 mg. Considering that the median body weight of subjects in the 130 mg group was 72 kg, the 130 mg ustekinumab dose was equivalent to approximately 2 mg / kg based on body weight. Therefore, on average, the ustekinumab exposure in the approximately 6 mg / kg group was approximately three times that of the 130 mg group. Consistent with this expectation, after a single IV administration of approximately 6 mg / kg or 130 mg of ustekinumab, the median serum ustekinumab concentration was approximately dose-proportional at all sampling time points throughout the 8 weeks. The median peak serum ustekinumab concentration observed 1 hour after the end of infusion at week 0 was 127.0 μg / mL and 43.16 μg / mL for the approximately 6 mg / kg group and the 130 mg group, respectively. At week 8, the time to evaluate the primary efficacy endpoint, the median serum ustekinumab concentration was 8.59 μg / mL and 2.51 μg / mL for the approximately 6 mg / kg group and the 130 mg group, respectively.

[0188] In subjects who received placebo IV in week 0 and did not show a clinical response at week 8, ustekinumab approximately 6 mg / kg was administered IV at week 8. Conversely, in subjects who received ustekinumab IV in week 0 and did not show a clinical response at week 8, ustekinumab 90 mg was administered via seroconcentrate (SC) at week 8. Even among subjects who received placebo IV in week 0 and then ustekinumab approximately 6 mg / kg IV at week 8, the median serum ustekinumab concentration at week 16 (8 weeks after ustekinumab IV administration) was slightly higher than that observed at week 8 (among subjects who received ustekinumab approximately 6 mg / kg IV at week 0, the concentrations were [10.51 μg / mL vs. 8.59 μg / mL, respectively]). Among the subjects who received ustekinumab 90 mg via saccharinic acid at week 8 (after the initial IV ustekinumab administration at week 0), the median serum ustekinumab concentration at week 16 was slightly higher in the subjects who received approximately 6 mg / kg of ustekinumab via IV at week 0 compared to the subjects who received 130 mg of ustekinumab at week 0 (1.92 μg / mL vs. 1.59 μg / mL, respectively).

[0189] Immunogenic results Of the 635 patients in the ustekinumab group who had suitable samples for evaluating antibodies against ustekinumab, 4 patients (0.6%) were positive for antibodies against ustekinumab throughout the 8-week period. Of these 4 patients, 2 patients (50%) were positive for NAb.

[0190] Of the 822 patients who received ustekinumab at any time throughout 16 weeks and had suitable samples for evaluation of anti-drug antibodies (ADA), 18 patients (2.2%) remained positive for antibodies against ustekinumab until the final safety evaluation visit. Of these, among those for whom NAb could be evaluated, 4 out of 15 patients (26.7%) remained positive for NAb until the final safety evaluation visit. In patients who received ustekinumab 90 mg via SC at week 8, the incidence of antibodies against ustekinumab throughout 16 weeks was numerically higher in the 130 mg IV → 90 mg SC group compared to the approximately 6 mg / kg IV → 90 mg SC group (4.5% [6 out of 132 patients] vs. 1.0% [1 out of 101 patients]).

[0191] Results of effectiveness Clinical remission at 8 weeks - World definition At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (15.5% and 15.6%, respectively) achieved clinical remission compared to the placebo group (5.3%; p<0.001 for both comparisons; Table 1).

[0192] [Table 1] N = number of subjects, CI = confidence interval

[0193] Clinical remission at 8 weeks - US definition At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (18.9% and 16.6%, respectively) achieved clinical remission compared to the placebo group (6.3%; p<0.001 for both comparisons; Table 2).

[0194] [Table 2] N = number of subjects, CI = confidence interval

[0195] Endoscopic healing at 8 weeks At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (27.0% and 26.3%, respectively) achieved endoscopic cure compared to the placebo group (13.8%; p<0.001 for both comparisons; Table 3).

[0196] [Table 3] N = number of subjects, CI = confidence interval

[0197] Clinical response at week 8 At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (61.8% and 51.3%, respectively) achieved a clinical response compared to the placebo group (31.3%; p<0.001 for both comparisons; Table 4).

[0198] [Table 4] N = number of subjects, CI = confidence interval

[0199] Change from baseline in the total IBDQ score at week 8. At baseline, the median IBDQ score was similar across all treatment groups. At week 8, the median improvement in IBDQ score from baseline was significantly greater in the approximately 6 mg / kg and 130 mg groups (31.0 and 31.5, respectively) compared to the placebo group (10.0; p<0.001 for both comparisons).

[0200] Clinical remission at 8 weeks When remission was assessed as clinical remission (world definition) by a rectal bleeding subscore of 0 at week 8, the proportion of subjects achieving this endpoint was nearly identical to the proportion observed based on the primary efficacy analysis (world definition). Approximately 15.2% and 15.3% of subjects in the 6 mg / kg and 130 mg groups achieved this endpoint, significantly higher proportions compared to the placebo group (5.3%; p<0.001 for both comparisons).

[0201] Symptomatic remission in the 8th week At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (44.7% and 41.3%, respectively) achieved symptomatic remission compared to the placebo group (22.6%; p<0.001 for both comparisons).

[0202] Histological healing at 8 weeks Histological healing was defined as the presence of 0–<5% neutrophils in the epithelium and the absence of crypt destruction, erosion, ulceration, or granulation tissue. At week 8, a significantly larger proportion of subjects in the 6 mg / kg and 130 mg groups (35.6% and 37.9%, respectively) achieved histological healing compared to the placebo group (21.9%; p<0.001 for both comparisons).

[0203] Change from baseline in the Mayo score at week 8. At baseline, the mean Mayo score was the same across all treatment groups (8.9 for all groups). At week 8, the mean reduction in Mayo score from baseline was significantly greater in the approximately 6 mg / kg and 130 mg groups (3.5 and 3.2, respectively) compared to the placebo group (1.8; p<0.001 for both comparisons).

[0204] Change from baseline in partial Mayo scores over 8 weeks. At baseline, the mean partial Mayo score was the same across all treatment groups (6.2 for all groups). As early as week 2 and throughout visits up to week 8, the mean decrease in partial Mayo score was significantly greater in the approximately 6 mg / kg and 130 mg groups compared to the placebo group. At week 2, the mean decrease in partial Mayo score from baseline was 1.6 and 1.5 in the approximately 6 mg / kg and 130 mg groups, respectively, compared to 1.0 in the placebo group (p<0.001 for both comparisons). At week 8, the mean decrease in partial Mayo score from baseline was 2.9 and 2.6 in the approximately 6 mg / kg and 130 mg groups, respectively, compared to 1.5 in the placebo group (p<0.001 for both comparisons).

[0205] UCEIS score in week 8 The UCEIS score provides a comprehensive assessment of endoscopic severity of ulcerative colitis (UC) based on mucosal vascular patterns, bleeding, and ulceration. The score ranges from 3 to 11, with higher scores indicating more severe disease as detected by endoscopy. The UCEIS score was assessed only during the central reading of the endoscopic video.

[0206] At baseline, mean UCEIS scores were similar across all treatment groups (7.6, 7.5, and 7.5 in the approximately 6 mg / kg, 130 mg, and placebo groups, respectively). At week 8, the mean reduction in UCEIS score from baseline was significantly greater in the approximately 6 mg / kg and 130 mg groups (1.3 and 1.1, respectively) compared to the placebo group (0.5) (p<0.001 for both comparisons).

[0207] At week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups (20.2% and 19.1%, respectively) had a UCEIS score of ≤4 compared to the 11.0% in the placebo group (p<0.001 and p=0.004, respectively). A UCEIS score of ≤4 is assumed to be associated with a Mayo endoscopic subscore of 0 or 1, which defined endoscopic cure as defined in this study.

[0208] Bristol Stool Consistency Scale Score The BSFS score at the time of visit was the average of the daily averages of the BSFS scores over the three days prior to the visit. The mean BSFS score at the time of visit was calculated using the same three days used to calculate the number of bowel movements and the rectal bleeding subscore of the Mayo score.

[0209] Approximately 40% (370 / 961 patients) of the randomized subjects had baseline BSFS scores. At baseline, 99.2% (367 / 370 patients) had a mean BSFS score of ≥3, and the majority of subjects (54.3%) had a mean BSFS score of ≥6, indicating diarrhea. As early as week 2, and continuing throughout visits up to week 8, the proportion of subjects with diarrhea (mean BSFS score ≥6) was lower in the approximately 6 mg / kg and 130 mg groups compared to the placebo group. At week 8, 22.8%, 21.1%, and 32.0% of subjects in the approximately 6 mg / kg, 130 mg, and placebo groups, respectively, had diarrhea (mean BSFS score ≥6). Furthermore, at week 8, the proportion of subjects with normal stool (≥3 and <5) was higher in the approximately 6 mg / kg group and the 130 mg group compared to the placebo group (48.3%, 48.9%, and 29.3%, respectively).

[0210] Normalization of C-reactive proteins C-reactive protein (CRP) is used as a marker of inflammation in individuals with IBD. In UC, high CRP is associated with severe clinical activity, high sedimentation rates, and active disease detected by colonoscopy. C-reactive protein is tested using validated, high-sensitivity CRP assays.

[0211] At baseline, the proportion of subjects with abnormal CRP levels (>3 mg / L) was similar across all treatment groups, with 59.2% of randomized subjects having abnormal CRP levels at baseline. As early as week 2, and continuing throughout visits up to week 8, a significantly larger proportion of subjects in the approximately 6 mg / kg and 130 mg groups, compared to the placebo group, achieved normalization of CRP levels (≤3 mg / L) among those with abnormal baseline levels. At week 8, 38.7% and 34.1% of subjects in the approximately 6 mg / kg and 130 mg groups, respectively, achieved normalization of CRP levels compared to 21.1% in the placebo group (p<0.001 for both comparisons).

[0212] Normalization of fecal lactoferrin levels At baseline, the proportion of subjects with abnormal fecal lactoferrin levels (>7.24 μg / g) was similar across all treatment groups, and overall, 90.0% of randomized subjects had abnormal fecal lactoferrin concentrations at baseline. At weeks 4 and 8, among subjects with abnormal baseline values, a significantly larger proportion in the approximately 6 mg / kg and 130 mg groups compared to the placebo group achieved normalization of fecal lactoferrin levels (≤7.24 μg / g). At week 8, 14.6% and 17.2% of subjects in the approximately 6 mg / kg and 130 mg groups, respectively, achieved normalization of fecal lactoferrin levels compared to 9.3% in the placebo group (p=0.042 and p=0.006, respectively, for the ustekinumab groups).

[0213] Normalization of fecal calprotectin levels At baseline, the proportion of subjects with abnormal fecal calprotectin levels (>250 mg / kg) was slightly higher in the approximately 6 mg / kg group (85.1%) compared to the placebo group (78.4%), and 82.5% of subjects in the 130 mg group had abnormal fecal calprotectin levels at baseline. At weeks 2 and 4, among subjects with abnormal values ​​at baseline, a significantly larger proportion in the approximately 6 mg / kg group and the 130 mg group achieved normalization of fecal capprotectin levels (≤250 mg / kg). At week 8, the proportion of subjects with normalized fecal calprotectin levels among those with abnormal fecal calprotectin levels at baseline was numerically higher in the ustekinumab approximately 6 mg / kg group and the 130 mg group (25.5% and 24.2%, respectively) compared to the placebo group (20.4%), although this was not statistically significant (p=0.148 and p=0.301 for the comparison between the two groups, respectively).

[0214] Example 2: Maintenance study of ustekinumab in the treatment of human ulcerative colitis methodology In this randomized treatment discontinuation maintenance trial, all enrolled subjects were respondents to the investigational drug administered in the induction trial. Primary (randomized) population: Subjects who showed a clinical response to IV ustekinumab after induction constituted the primary population in the maintenance trial. This population included: subjects who were randomized at week 0 of the induction trial to receive ustekinumab (i.e., 130 mg IV or approximately 6 mg / kg IV) and showed a clinical response at week 8 of induction; and subjects who were randomized at week 0 of the induction trial to receive placebo, did not show a clinical response at week 8 of induction, but showed a clinical response at week 16 of induction after receiving a dose of IV ustekinumab (approximately 6 mg / kg) at week 8 of induction (placebo → ustekinumab approximately 6 mg / kg IV). These subjects were randomized in a 1:1:1 ratio at week 0 of the maintenance phase to receive ustekinumab 90 mg SC every 8 weeks (q8w), ustekinumab 90 mg SC every 12 weeks (q12w), and placebo SC. Non-randomized population: Additional subjects entering the maintenance trial were not randomized in the primary population and received maintenance dosing as follows: Ustekinumab delayed responseers (i.e., subjects who did not show a clinical response to IV ustekinumab at week 8 of the induction phase but showed a clinical response at week 16 of the induction phase after administration of ustekinumab 90 mg SC at week 8 of the induction phase) received ustekinumab 90 mg SC q8w, and placebo responseers (i.e., subjects who showed a clinical response to placebo IV induction) received placebo SC. Non-randomized subjects were followed for both efficacy and safety but were not included in important efficacy analyses.

[0215] All subjects received their assigned dose of the SC study drug at their visit during week 0 of the maintenance phase. Thereafter, to maintain blinding, all subjects received the study drug at every scheduled visit. Subjects were assessed for clinical flare at each visit, and rescue medication was administered if a decline in clinical response was observed. The main portion of the maintenance study ran until week 44, with the long-term study extension continuing until week 220.

[0216] Number of subjects (planned and analyzed): 783 patients who completed the induction trial and demonstrated a clinical response to the induction drug were enrolled in this maintenance trial. The number of subjects in each treatment group at week 0 of the maintenance phase was as follows: ● Randomized (primary) population (523 cases [327 cases planned]): -176 patients were randomized to ustekinumab 90 mg SC q8w. -172 patients were randomized to ustekinumab 90 mg SC q12w. -175 cases were randomized to placebo SC. ●Non-randomized group (260 cases): -157 patients who were delayed responders to ustekinumab (i.e., those who did not show a clinical response to ustekinumab at week 8 of the induction period but showed a clinical response at week 16 of the induction period) were administered ustekinumab 90 mg SC q8w. -103 patients who clinically responded to IV induction of placebo (placebo-initiated responders) received placebo via SC administration.

[0217] Diagnostic and Key Criteria for Inclusion All subjects enrolled in this randomized treatment discontinuation maintenance trial were those with moderate to severe active ulcerative colitis who had an inadequate response to or poor tolerance to conventional therapy (i.e., corticosteroids or immunomodulators) or biological therapy (i.e., TNF antagonists and / or vedolizumab) and who demonstrated a clinical response to the study drug during the induction trial. This included subjects who demonstrated a clinical response to IV ustekinumab, a clinical response to IV placebo, or a delayed clinical response to ustekinumab and who did not undergo any drug changes prohibited in the study protocol during the induction trial.

[0218] Evaluation criteria: ●Pharmacokinetics (PK): Serum ustekinumab concentration ●Immunogenicity: Antibodies against ustekinumab ●Pharmacodynamics (PD) / Biomarkers: Serum biomarkers; fecal microbiome; RNA expression and histological evaluation of disease activity and healing in mucosal biopsies ● Genetics and Epigenetics: Whole blood deoxyribonucleic acid (DNA) ●Efficacy: Mayo score and partial Mayo score, UC Endoscopic Severity Assessment Index (UCEIS), CRP, fecal lactoferrin, and fecal calprotectin ● Health-related quality of life: Inflammatory Bowel Disease Questionnaire (IBDQ), 36-item Short-Form Health Survey (SF-36), and EuroQoL-5D Health Questionnaire (EQ-5D) ● Health Economics: Hospitalizations and surgeries related to UC disease; Visual Analog Scale (VAS), and Work Productivity and Activity Impairment Questionnaire - General Health (WPAI-GH) ●Safety: Adverse events (AEs), serious adverse events (SAEs), infections, injection site reactions, allergic reactions, hematological and chemical parameters, vital signs, physical examination, and early detection of tuberculosis.

[0219] endpoint ● The primary endpoint was clinical remission at week 44. The definition of clinical remission (and the study procedure) differed between the U.S. and international applications to adapt to the preferred global and U.S. definitions of clinical remission. Each definition of clinical remission was applied to all subjects in the primary efficacy analysis population. - The global definition of the primary endpoint for clinical remission is a Mayo score of 2 or less, with no individual subscores exceeding 1. -The U.S. definition of clinical remission was defined as an absolute fecal count of 3 or less, a Mayo rectal bleeding subscore of 0, and a Mayo endoscopic subscore of 0 or 1. ●The primary secondary endpoints, listed in the order they were tested, were as follows: - Maintaining clinical response up to week 44 - Endoscopic healing at 44 weeks - Clinical remission and no concomitant corticosteroid administration at week 44 (corticosteroid-free clinical remission) - Maintenance of clinical remission up to week 44 among subjects who achieved clinical remission at baseline during the maintenance phase.

[0220] For the third and fourth primary secondary endpoints, we used the global definition of clinical remission to support applications in countries outside the U.S., and the U.S. definition of clinical remission to support applications in the U.S.

[0221] Demographic and baseline disease characteristics were compiled based on 961 cases in the primary efficacy analysis population.

[0222] Except for a fourth primary secondary endpoint related to the maintenance of clinical remission, analyses of multiple controlled endpoints were performed using the Cochrane-Mantel-Haenszel (CMH) chi-squared test, stratified by clinical remission (world definition) status at baseline during the maintenance phase (yes / no as determined by the IWRS) and induction treatment (placebo IV [I-0] → ustekinumab approx. 6 mg / kg IV [I-8], ustekinumab 130 mg IV [I-0], or ustekinumab approx. 6 mg / kg IV [I-0]). For the fourth primary secondary endpoint (maintenance of clinical remission), the CMH chi-squared test stratified by induction dose was used.

[0223] Multiple global and US-specific trial procedures were pre-specified to control for the overall Type 1 error rate at the 0.05 level across the multiplicity-controlled endpoints in this trial (Section 3.11.2.7.3). All statistical tests were performed at a two-tailed 0.05 significance level. Nominal p-values ​​are presented.

[0224] Safety was assessed collectively by evaluating the frequency and type of adverse events (AEs), laboratory parameters (hematology and chemistry), and vital sign parameters that occurred during treatment. Safety summaries are provided separately for randomized subjects, non-randomized subjects, and all treated subjects. The presentation of safety data focuses on the randomized population.

[0225] result Test population A total of 783 patients who completed the induction trial and demonstrated a clinical response to the induction drug were enrolled in this maintenance trial. Of these, 523 belonged to the primary population targeted for the maintenance trial and were randomized to receive ustekinumab or placebo SC at week 0 of the maintenance phase (176, 172, and 175 patients in the ustekinumab 90 mg SC q8w, ustekinumab 90 mg SC q12w, and placebo groups, respectively). The remaining 250 patients belonged to a non-randomized population, including 157 delayed ustekinumab induction responders (who received ustekinumab 90 mg SC q8w) and 103 placebo induction responders (who received placebo). All enrolled subjects assigned treatment at baseline in the maintenance phase received the corresponding investigational drug at that time.

[0226] Prior to week 40 (visit for the final dose of the maintenance trial), 85 subjects (16.3%) in the primary population discontinued the study drug. The proportion of subjects discontinuing the study drug was higher in the placebo group (24.6%) than in the ustekinumab q8w and q12w groups (10.2% and 14.0%, respectively). The most common reasons for discontinuation were lack of efficacy due to worsening of UC and adverse events. Prior to week 44, 29 subjects (5.5%) in the primary population terminated participation in the trial, with the most common reason for termination being withdrawal of consent.

[0227] Baseline clinical disease characteristics represented a population of subjects with moderate to severe active UC refractory to available therapies and were generally well-balanced across the three treatment groups. The median duration of disease was 6.05 years, the median baseline Mayo score was 9.0, and 86.9% and 13.1% had moderate and severe UC, respectively. At induction baseline, 52.2% of subjects in the primary population of the maintenance study were taking corticosteroids, 26.6% were taking immunomodulators, and 70.7% were taking aminosalicylates. The majority of subjects (93.5%) had an inadequate response to or poor tolerance of corticosteroids and / or 6-MP / AZA, or demonstrated corticosteroid dependence at induction baseline. Overall, in the primary population, 47.6% of subjects had a history of documented biological agent refractory, while 52.4% did not. Furthermore, 47.2% were unsuccessful with at least one anti-TNF agent, 13.4% were unsuccessful with both anti-TNF agents and vedolizumab, 49.3% were inexperienced with biological therapies, and two patients were unresponsive to vedolizumab alone.

[0228] Results of effectiveness Ustekinumab maintenance therapy demonstrated efficacy in a population of patients with moderate to severe active UC who had previously failed or had poor tolerance to conventional or biological therapies including TNF antagonists and / or vedolizumab, and who showed a clinical response at 8 weeks after receiving a single induction dose of ustekinumab IV.

[0229] Based on pre-specified global and US-only trial procedures, statistical significance can be claimed for both ustekinumab regimens (90 mg at q8w and 90 mg at q12w) for the primary endpoint of clinical remission at 44 weeks and three major secondary endpoints: maintenance of clinical response throughout 44 weeks, endoscopic cure at 44 weeks, and corticosteroid-free clinical remission at 44 weeks. In addition, statistical significance can be claimed for maintenance of clinical remission throughout 44 weeks (among subjects who achieved clinical remission at baseline during the maintenance period) for both ustekinumab doses based on US-only trial procedures and the ustekinumab q12w regimen based on global trials.

[0230] ●Clinical efficacy in the main population (i.e., subjects with a clinical response 8 weeks after receiving IV induction therapy with ustekinumab) - Primary endpoint: Clinical remission ○The proportion of subjects in clinical remission (based on the world definition) at week 44 was significantly higher in the ustekinumab q8w group and the ustekinumab q12w group (43.8% and 38.4%, respectively) compared to the placebo group (24.0%) (p<0.001 and p=0.002, respectively). ○The proportion of subjects in clinical remission (based on the US-specific definition) at week 44 was significantly higher in the ustekinumab q8w group and the ustekinumab q12w group (42.6% and 39.5%, respectively) compared to the placebo group (24.6%) (p<0.001 and p=0.002, respectively). The effect of ustekinumab on achieving clinical remission (based on both the global and limited US definitions) was generally consistent across subgroups (including subjects who were refractory to biological agents and those who were not, as well as subjects who received concomitant immunomodulators or corticosteroids at baseline during the induction phase and those who did not) and was robust to pre-defined changes in data processing rules. - Primary secondary endpoints: maintenance of clinical response, endoscopic cure, corticosteroid-free clinical remission, and maintenance of clinical remission. The proportion of patients who maintained a clinical response throughout 44 weeks, achieved endoscopic cure, and achieved corticosteroid-free remission (applying both the global and US-specific definitions of clinical remission) was significantly higher in the ustekinumab q8w and q12w groups compared to the placebo group (p<0.01). ○The proportion of subjects who maintained clinical remission among those who achieved clinical remission at baseline during the maintenance phase was numerically higher in both the ustekinumab q8w and q12w groups compared to the placebo group (applying both the global and US-specific definitions of clinical remission). Statistical significance (p<0.01) was achieved for both the q8w and q12w groups compared to placebo when using the US-specific definition of clinical remission, but statistical significance was achieved only for the q12w group compared to placebo when using the global definition of clinical remission (p<0.01). - Other histological, mucosal, clinical, and endoscopic endpoints The analyses summarized below were not controlled for multiplicity. Statistical significance is described based on nominal p-values. ○The proportion of subjects who achieved histological healing at week 44 (i.e., <5% neutrophil infiltration in crypts, no crypt destruction, and no erosion, ulceration, or granulation tissue) was significantly greater in the ustekinumab q8w and q12w groups compared to the placebo group (p<0.001). ○The proportion of subjects who achieved mucosal healing (a combination of endoscopic and histological healing) at week 44 was significantly higher in the ustekinumab q8w and q12w groups compared to the placebo group (p<0.01). Applying both the global and US-specific definitions of clinical remission, the proportion of patients achieving corticosteroid-free remission for at least 90 days prior to 44 weeks was significantly higher in the ustekinumab q8w and q12w groups compared to the placebo group (p<0.01). Furthermore, among patients receiving corticosteroids at baseline during the maintenance phase, a significantly larger proportion (p<0.05) of patients in the ustekinumab q8w and q12w groups showed clinical remission and were not receiving concomitant corticosteroids for at least 90 days prior to 44 weeks, compared to the placebo group. The efficacy of ustekinumab maintenance therapy was also demonstrated in clinical outcomes, measured by sustained improvement in partial Mayo scores, maintenance of symptomatic remission, and maintenance of endoscopic healing. Further evidence of the efficacy of ustekinumab maintenance therapy was observed in longitudinal partial Mayo remission and symptomatic remission, as well as in symptom control (bowel movement frequency and rectal bleeding). - Inflammatory biomarkers ○Throughout the period up to week 44, the ustekinumab group maintained the CRP, fecal lactoferrin, and fecal calprotectin levels observed at the maintenance baseline, while in the placebo group, median CRP, fecal lactoferrin, and fecal calprotectin levels worsened (increased). ○At week 44, the proportion of subjects with normalized CRP, fecal calprotectin, and fecal lactoferrin was generally significantly higher in the ustekinumab q8w and q12w groups compared to the placebo group. - Clinical endpoints by state of biological failure Regarding subjects with and without a history of resistance to biological agents, the proportion of subjects achieving the primary endpoint, major secondary endpoints, and mucosal healing was generally higher in the ustekinumab q8w and q12w groups compared to the placebo group. In some cases, when the efficacy of administration was similar in the biological agent-responsive population and the biological agent-ineffective population, a consistent trend across endpoints was observed in the biological agent-ineffective population: the efficacy of ustekinumab in the q8w group was greater than that of ustekinumab in the q12w group. This trend was not observed in the biological agent-responsive population. - Efficacy based on subgroups of inflammatory biomarkers In subjects with a higher inflammatory load (elevated CRP and / or fecal inflammatory markers) at baseline in either the induction or maintenance phase, both doses generally showed efficacy compared to placebo. However, the efficacy of ustekinumab q8w appeared to be better than that of ustekinumab q12w across the range of clinical endpoints. Nevertheless, in subjects with a low baseline inflammatory load, both the ustekinumab q8w and q12w groups showed similar efficacy for the endpoints. - Health-related quality of life ○By week 44, participants in the ustekinumab q8w and q12w groups were generally able to maintain health-related quality of life improvements compared to the placebo group, as assessed using the IBDQ, SF36, and EQ5D assessment methods. - Results of ustekinumab 90 mg administration over 8 weeks and ustekinumab 90 mg administration over 12 weeks Both the ustekinumab q8w and q12w groups generally showed similar efficacy for the primary and primary secondary endpoints, although q8w was slightly better than q12w based on the following more objective and rigorous efficacy measures. ◆ Endoscopic and mucosal healing at 44 weeks ◆Long-term partial Mayo remission at week 44 ◆Among subjects receiving corticosteroids at baseline during the maintenance phase, clinical remission and corticosteroid elimination for at least 90 days prior to 44 weeks. ○Furthermore, when efficacy was tested over time (for the following endpoints), the q8w group showed greater efficacy than the q12w group: ◆Mayo stool frequency and rectal bleeding subscores indicating inactive or mild disease (i.e., subscore of 0 or 1), and ≤3 absolute stool counts over the period up to week 44. ◆Partial Mayo remission and symptomatic remission over the period up to week 44. ◆Median change over time from baseline in fecal lactoferrin and calprotectin concentrations over the period up to week 44.

[0231] ●Efficacy in ustekinumab-introduction delayed responders Subjects who were delayed responders to ustekinumab induction therapy were able to maintain clinical response and achieve clinical remission, endoscopic healing, histological healing, and mucosal healing (combination of endoscopic and histological healing) while receiving ustekinumab 90 mg q8w.

[0232] ●Efficacy and pharmacokinetics / immunogenicity -Generally, during maintenance, a clear association was observed between serum ustekinumab concentration and the clinical efficacy outcomes of clinical remission and endoscopic healing. In addition, lower levels of inflammation were observed in subjects with higher serum ustekinumab concentrations when measured by CRP. -Among subjects receiving maintenance ustekinumab, the expression of antibodies to ustekinumab did not appear to affect clinical efficacy measured by multiple endpoints such as clinical remission, endoscopic healing, clinical response, and change from maintenance baseline in Mayo score, although the interpretation of the data was limited by the small sample size.

[0233] Pharmacokinetic and immunogenicity results ● Following maintenance administration of ustekinumab 90 mg via SC at q8w or q12w, subjects reached a steady state approximately 8 or 12 weeks after initiating a maintenance regimen of ustekinumab 90 mg SC q8w or ustekinumab 90 mg SC q12w, respectively. The median steady-state trough serum ustekinumab concentration over time was approximately three times higher in the ustekinumab q8w group (2.69 μg / mL to 3.09 μg / mL) than in the q12w group (0.92 μg / mL to 1.19 μg / mL). ● Following maintenance regimens of ustekinumab 90 mg administered via q8w or q12w, serum ustekinumab concentrations persisted until week 44 in almost all subjects. The proportion of subjects with undetectable trough concentrations over time was lower in the 90 mg q8w group (0.7%–2.4%) compared to the 90 mg q12w group (4.9%–7.1%). Median ustekinumab concentrations in the placebo group fell below detectable levels by week 16. ●The effect of different ustekinumab IV induction doses on maintenance serum ustekinumab concentrations continued to decrease over time, as expected. ●The median trough serum ustekinumab concentration tended to be lower in subjects with higher body weight. ●In the ustekinumab delayed response group, non-randomized subjects tended to have lower serum ustekinumab concentrations over time after SC administration of the same ustekinumab regimen of 90 mg for q8 weeks, compared to randomized subjects in the ustekinumab q8w group. ●Of the 680 subjects for whom appropriate samples were available to evaluate antibodies against ustekinumab, 39 subjects (5.7%) tested positive for antibodies against ustekinumab throughout 52 weeks of administration, with the majority having antibody titers of ≤1:800. In this maintenance study, of the 39 subjects who tested positive for antibodies against ustekinumab, 11 subjects (28.2%) tested positive for neutralizing antibodies. ●In all randomized treatment groups, the median serum ustekinumab concentration was lower over time in subjects who were positive for antibodies to ustekinumab compared to subjects who were negative for antibodies to ustekinumab.

[0234] Safety results Subcutaneous maintenance regimens of ustekinumab 90 mg administered at q12W or q8W up to week 44 were generally well-tolerated and consistent with the known safety profile of ustekinumab. ● AEs were reported in 77.3%, 69.2%, and 78.9% of the subjects in the ustekinumab q8w group, ustekinumab q12w group, and placebo group, respectively. - Reasonably causal adverse events (AEs) were reported in 26.1%, 17.4%, and 28.6% of subjects in the ustekinumab q8w group, the ustekinumab q12w group, and the placebo group, respectively. ● Infections (identified by the investigator) were reported in 48.9%, 33.7%, and 46.3% of the subjects in the ustekinumab q8w group, ustekinumab q12w group, and placebo group, respectively. Infections requiring oral or parenteral antibiotic treatment were reported in 22.7%, 15.7%, and 19.4% of the subjects in the ustekinumab q8w group, ustekinumab q12w group, and placebo group, respectively. ●Serious infections were infrequent among the randomized subjects, reported in 1.7%, 3.5%, and 2.3% of the ustekinumab q8w, ustekinumab q12w, and placebo groups, respectively. Opportunistic infections were identified in 3 subjects (all in the randomized population). Cytomegalovirus colitis was diagnosed in 2 cases in the ustekinumab q12w group, one of which was diagnosed as a moderate concurrent AE of ocular and oral herpes. No cases of active TB were reported among the ustekinumab recipients throughout 44 weeks. ● The proportion of randomized subjects experiencing adverse events (AEs) that led to discontinuation of the study drug was higher in the placebo group than in the q12w and q8w groups, and the most frequent AE leading to discontinuation in the placebo group was exacerbation of ulcerative colitis (UC). ●Among all recipients, including those with delayed responses to ustekinumab, the overall safety profile was consistent with that observed in the randomized population. ●A death was reported in one patient who was a delayed responder to ustekinumab induction and was receiving ustekinumab at q8w. The cause of death was acute respiratory failure that occurred during thyroid surgery for multinodular goiter. ● Of all patients treated, two (one belonging to the ustekinumab induction-delayed response group [ustekinumab administered at q8w], and the other randomized to a placebo group receiving IV ustekinumab I during induction) reported serious major adverse cardiovascular events, and both events were associated with perioperative complications. ●Among all patients treated, six cases of malignant disease were reported (five patients treated with ustekinumab and one patient treated with placebo only). - Three patients treated with ustekinumab reported non-melanoma skin cancer (NMSC), all of whom had a history of either azathioprine or 6-MP treatment, and two were receiving concomitant immunomodulatory therapy at the time of diagnosis. Two of the ustekinumab recipients were reported to have solid tumors; one had papillary renal cell carcinoma (q12w) and the other had colon cancer (q8w). Both tumors were detected early during the subjects' participation in this maintenance study. ● No cases of anaphylactic or delayed-type hypersensitivity reactions were identified among patients treated with ustekinumab. ● There was no significant difference in the proportion of subjects with maximum toxicity grade chemical and hematological laboratory values ​​of ≥1 after baseline between the placebo group and the respective ustekinumab groups. Grade 3 and grade 4 chemical and hematological laboratory values ​​were infrequent.

[0235] Results of healthcare economics and utilization of medical resources ●By week 44, patients in the ustekinumab group receiving concomitant therapy had significantly fewer hospitalizations or surgeries related to UC disease compared to the placebo group. ●At week 44, changes from maintenance baseline in the Visual Analog Score (VAS) of productivity demonstrated improvement in the ustekinumab group and deterioration in the placebo group. ●At week 44, the percentages in each of the four WPAI-GH domains were maintained from baseline in the ustekinumab treatment group, and additional improvements were observed in the health-related work-related impairment rate, health-related overall work impairment rate, and health-related activity impairment rate in the ustekinumab q8w group. In the placebo group, the percentages in all four WPAI-GH domains worsened (i.e., increased).

[0236] conclusion ● The ustekinumab maintenance study consistently provided conclusive evidence that both administration regimens of ustekinumab 90 mg administered via sustained saturation (SC) at q12w and q8w were effective in adult patients with moderate to severe active UC who had responded to a single IV ustekinumab induction dose. - The efficacy of ustekinumab was observed in patients who were unresponsive to biological agents, as well as in patients who were unsuccessful with conventional therapy but not with biological therapy (i.e., biological agent-naive). Notably, while both doses of ustekinumab were effective, the q8w dosing regimen showed slightly better efficacy across several objective and / or more stringent endpoints (e.g., endoscopic cure and long-term partial Mayo remission), as well as over time analyses of symptomatic and partial Mayo remission. ● Maintenance doses of ustekinumab in the SC regimens of 90 mg for q12w and 90 mg for q8w were generally well-tolerated over 44 weeks in a population of adults with moderate to severe ulcerative colitis. ●The safety and efficacy data from this trial support a favorable benefit / risk profile for ustekinumab SC maintenance therapy.

[0237] Example 3: Long-term continuation of ustekinumab in a maintenance study for the treatment of ulcerative colitis. Clinical trial protocol CNTO1275UCO3001; Long-term continuation of administration in a Phase 3 maintenance study Clinical trial protocol number: CNTO1275UCO3001 Study Name: Phase 3, randomized, double-blind, placebo-controlled, parallel-group, multicenter study to evaluate the safety and efficacy of ustekinumab induction and maintenance therapy in patients with moderate to severe active ulcerative colitis. Test name: UNIFI EudraCT number: 2014-005606-38 NCT ID: NCT02407236 Clinical registration number: CR106920 Principal Investigator: Bruce Sands, MD, (Division of Gastroenterology, Icahn School of Medicine at Mount Sinai; New York, NY, USA). Test centers: 201 locations across Asia, Eastern Europe, North America, Western Europe, Israel, Australia, and New Zealand. Publications (References): Sands BE, Sandborn WJ, Panaccione R, et al. Ustekinumab as Induction and Maintenance Therapy for Ulcerative Colitis. N Engl J Med. 2019;381(13):1201-1214. Trial period: August 19, 2015 (the date the first subject signed informed consent) to August 12, 2019 (the last observation date the last subject was recorded as part of the database) Phase 3 of development: Objective: The objective of the long-term study extension (LTE) trial was to evaluate the efficacy, safety, pharmacokinetics (PK), and immunogenicity of ustekinumab when administered for an additional year in subjects with moderate to severe active ulcerative colitis (UC) who had completed a 44-week maintenance trial and in subjects determined by the investigator to benefit from continued treatment. Methodology: Safety and efficacy evaluations were completed at week 44 of the maintenance trial. Subjects determined by the investigator to benefit from continued treatment were offered the opportunity to participate in LTE for an additional three years. Randomized population: The primary (randomized) population in the maintenance trial consisted of subjects who had demonstrated a clinical response to IV ustekinumab after the induction phase. Subjects were randomized at maintenance baseline to receive placebo SC, ustekinumab 90 mg SC every 12 weeks (q12w), or ustekinumab 90 mg SC every 8 weeks (q8w). Non-randomized population: Additional subjects entering the maintenance trial included: subjects who received placebo SC during maintenance and had demonstrated a clinical response during the placebo IV induction phase (i.e., placebo induction responders), and subjects with a delayed ustekinumab response (i.e., subjects who did not demonstrate a clinical response to ustekinumab at week 8 of the induction phase but did demonstrate a clinical response at week 16 of the induction phase after receiving ustekinumab SC at week 8 of the induction phase), who received ustekinumab 90 mg SC q8w during maintenance (i.e., delayed ustekinumab responders).

[0238] During the LTE period, participants continued the same dosing regimen as at week 44 of the maintenance trial (either placebo, ustekinumab 90 mg SC q12w, or ustekinumab 90 mg SC q8w), with the first dose in the LTE administered at week 48. During the LTE period, all participants needed to be evaluated for worsening UC disease activity based on the clinical judgment of the principal investigator. Among the primary population (participants randomized at week 0 of the maintenance phase), those with worsening UC disease activity received a single dose adjustment as follows: placebo SC → ustekinumab 90 mg SC q8w; ustekinumab 90 mg SC q12w → ustekinumab 90 mg SC q8w; ustekinumab 90 mg SC q8w → continued ustekinumab 90 mg SC q8w (sham dose adjustment). Participants' first visit to the hospital for dose adjustment consideration was at week 56. The target group was those who could adjust their dosage once during the LTE period.

[0239] During the LTE period, the blinded trial was maintained until the end of the maintenance study, which included the completion of the 44-week visit evaluation and the completion of the 44-week analysis. Therefore, participants continued to receive the study drug at their monthly visits until then. After the trial was unblinded at the clinical trial sites, participants who received placebo discontinued participation in the trial, while those who received ustekinumab continued to receive ustekinumab, but their trial visits were scheduled according to their administration regimen (either q8w or q12w, as appropriate for their regimen).

[0240] Number of subjects (planned and analyzed): 588 patients were included in the LTE (Low-Term Inhibitory) regimen if safety and efficacy evaluations were completed at week 44 and the principal investigator determined that continued administration would be beneficial.

[0241] ●The following 399 patients were randomized at maintenance baseline and received treatment during the LTE period: - Placebo SC: 115 cases - Ustekinumab 90mg SC q12w: 141 cases - Ustekinumab 90mg SC q8w: 143 cases Of the randomized population, a total of 32.6% (130 patients) underwent the following dose adjustments during the LTE period: - Of those randomized to placebo, 46.1% (53 patients) received dose adjustments to the ustekinumab 90 mg SC q8w regimen. Of the patients randomized to ustekinumab 90 mg SC q12w, 28.4% (40 patients) were dose-adjusted to the ustekinumab 90 mg SC q8w regimen. - Of the patients randomized to ustekinumab 90 mg SC q8w, 25.9% (37 patients) underwent sham dose adjustment (continuation of the same administration regimen).

[0242] ●The non-randomized group of 189 patients who received treatment during the maintenance phase in the LTE period is as follows: - Placebo SC: 73 subjects who showed a clinical response to placebo IV induction (placebo IV responseers) continued to receive placebo SC during the maintenance period and LTE period until blinding of the study was lifted and the study was discontinued. -Ustekinumab 90 mg q8w: 116 patients with delayed ustekinumab induction response (i.e., those who did not show a clinical response to ustekinumab at week 8 of the induction phase, but showed a clinical response at week 16 of the induction phase after receiving ustekinumab via SC at week 8 of the induction phase) were continuously administered ustekinumab 90 mg SC q8w during the maintenance period and up to LTE.

[0243] Diagnostic and main criteria for inclusion: At week 44 of the maintenance study, safety and efficacy evaluations were completed, and subjects who the principal investigator determined would benefit from continued administration were given the opportunity to participate in the LTE for an additional 3 years.

[0244] Test specimen, dosage and administration method, batch number: Ustekinumab was supplied as a sterile liquid for SC injection in single-use prefilled syringes (PFS). Each single-use PFS contained 90 mg (1.0 mL filled liquid; bulk lot numbers 14L012, 15K142, 16B012, 16H032, 16L012, 17B042, 17J012, 18B092, and FJZ02) of ustekinumab in an aqueous medium of L-histidine, L-histidine hydrochloride monohydrate, sucrose, and polysorbate 80 at pH 6.0. No preservatives were present.

[0245] Reference therapy, dosage and administration method, batch number: Placebo was supplied as sterile liquid for SC injection in single-use PFS (bulk lot numbers 15L042, 16L022, 17F042, EJSSL) in 1.0 mL fill volumes. Each PFS contained L-histidine, sucrose, and polysorbate 80 at pH 6.0. Placebo administrations had a similar appearance to each ustekinumab administration.

[0246] Duration of administration: The main part of the maintenance study was conducted until week 44, and LTE continued until week 220. The duration of administration in this initial part of LTE was 52 weeks (from maintenance week 44 to week 96).

[0247] Test evaluation: ●Pharmacokinetics: Serum ustekinumab concentration ●Immunogenicity: Antibodies against ustekinumab ●Efficacy: Partial Mayo score, C-reactive protein (CRP), fecal lactoferrin, fecal calprotectin, and use of corticosteroids ● Health-related quality of life: Inflammatory Bowel Disease Questionnaire (IBDQ), 36-item Short-Form Health Survey (SF-36) ● Health Economics: Hospitalizations and surgeries related to UC disease; Visual Analog Scale (VAS) for Productivity, and Work Productivity and Activity Impairment Questionnaire - General Health (WPAI-GH) ●Safety: AEs, serious adverse events (SAEs), infections, injection site reactions, allergic reactions, hematological and chemical parameters, vital signs, physical examination, and early detection of tuberculosis (TB).

[0248] statistical methods The primary objective of the efficacy analysis was to assess the maintenance of clinical utility from the end of the main trial (week 44) to week 92. Demographic characteristics, baseline disease characteristics, PK, immunogenicity, efficacy, and safety were analyzed for subjects receiving LTE (including both randomized and non-randomized subjects). Continuous variables were summarized using descriptive statistics (e.g., mean, median, standard deviation, interquartile range, minimum, and maximum). Categorical variables were summarized using counts and proportions. Statistical comparisons between treatment groups were not performed.

[0249] result This primarily aggregates data from weeks 44 to 96 of LTE data.

[0250] Test population At week 44, safety and efficacy evaluations were completed, and a total of 588 patients who, in the judgment of the principal investigator, were deemed to benefit from continued administration were included in the LTE (Long-Term Recovery) regimen and administered with the same dosing regimen as at week 44 of the maintenance phase. Of these, 399 patients were in the randomized maintenance population (115, 141, and 143 patients in the placebo, ustekinumab 90 mg SC q12w, and ustekinumab 90 mg SC q8w groups, respectively). The remaining 189 patients were in the non-randomized population, including 73 placebo-initiated respondents (placebo administration) and 116 delayed ustekinumab-initiated respondents (ustekinumab 90 mg SC q8w).

[0251] By week 96, 71 patients (17.8%) from the randomized population discontinued the study drug. In the ustekinumab combination group, the most common reasons for discontinuation were adverse events due to worsening UC (2.5% [7 patients]) and other reasons (2.5% [7 patients]; most reported as withdrawal of consent). Among the non-randomized group, 5 patients (4.3%) from the ustekinumab delayed response group discontinued the study drug. The most common reason for discontinuation was "other" (1.7% [2 patients]; both reported as withdrawal of consent).

[0252] The clinical disease characteristics at week 44 of randomized subjects receiving LTE were generally similar in the ustekinumab q12w and q8w groups, with numerically higher (e.g., Mayo score, CRP concentration) or lower (e.g., subjects in remission) values ​​in the placebo group, indicating greater disease activity in the placebo group. The clinical disease characteristics at week 44 of the ustekinumab delayed response group (ustekinumab administered at q8w during the LTE period) showed higher disease activity compared to the clinical disease characteristics of randomized subjects in the ustekinumab q8w group (e.g., fewer subjects in remission at the clinical efficacy endpoint, higher levels of inflammatory biomarkers, etc.).

[0253] The majority (93.7%) of patients who received LTE and were randomized during the maintenance phase either had an inadequate response to or poor tolerance of corticosteroids and / or 6-mercaptopurine / azathioprine at induction baseline, or exhibited corticosteroid dependence. Of the patients randomized during the maintenance phase after receiving LTE, 55.9% had no history of biological agent failure at induction baseline (53.1% were biological agent-naive, and 2.8% had no record of biological agent failure). A total of 44.1% of the randomized patients had a history of biological agent failure. The proportion of patients with a history of biological agent failure was lower in the ustekinumab q12w group (37.6%) compared to the ustekinumab q8w group (49.7%). The medical history of response to and tolerance of UC medications, as well as the medical history of UC medications, in the ustekinumab delayed response group who received treatment during the LTE period were generally consistent with that of the randomized population from the ustekinumab q8w group.

[0254] Pharmacokinetic and immunogenicity results ● After administration of ustekinumab 90 mg SC q8w or q12w during the LTE period, sustained concentrations of ustekinumab were observed up to week 92, and these concentrations were generally consistent with the serum ustekinumab concentrations observed in these treatment groups during the maintenance study. ● The expression rate of antibodies against ustekinumab remained low until week 96 of the LTE. -Of the 400 patients who received ustekinumab in both the induction and maintenance phases up to week 96 of the LTE, 22 patients (5.5%) were antibody-positive for ustekinumab up to week 96, and the antibody titer of most patients was 1:800 or less. -Of the 515 patients who received at least one dose of ustekinumab during the induction or maintenance phase up to week 96 of the LTE study, 34 patients (6.6%) were antibody-positive for ustekinumab by week 96 of this study, and the antibody titer of most patients was 1:800 or less. ○In this maintenance study, the expression rate of antibodies against ustekinumab appeared to be higher in subjects who were randomized to placebo (subjects who received a single initial infusion of ustekinumab during the induction phase), or in subjects who required dose adjustment from placebo or ustekinumab q12w during the LTE period. Of the 34 patients who received the treatment and tested positive for antibodies against ustekinumab, 8 patients (23.5%) tested positive for neutralizing antibodies.

[0255] Results of effectiveness The objective of the efficacy analysis in the Longest Live (LTE) was to evaluate the maintenance of clinical utility from the end of the primary trial (week 44) to week 92. It is important to note that participants entered the LTE based on the investigator's judgment that continued administration would provide a benefit. The placebo group represents a subgroup of UC patients who were either long-term responders to ustekinumab induction therapy (i.e., re-randomized during the placebo maintenance phase) or placebo induction responders with a long disease latency period. For these reasons, and because the placebo group was to discontinue participation in the trial after deblinding, direct comparison of the results between the treatment groups was considered to be confounding. Therefore, statistical comparisons were not performed.

[0256] Randomized subjects who received LTE ●From week 44 to week 92, the rates of symptomatic remission and partial Mayo remission in the randomized population of the ustekinumab q12w and q8w groups were maintained. -Similar sustained efficacy was observed in the group previously untreated with biological agents, the group that responded to biological agents, and the group that did not respond to biological agents. ●When ustekinumab administration was continued in the LTE study, the subjects were able to achieve symptomatic remission and partial Mayo remission at week 92 in the absence of corticosteroids. ●Continuing administration of ustekinumab allowed the patient to eliminate corticosteroids. ●If ustekinumab administration is continued from week 44 to week 92: - The reduction in partial Mayo scores observed at baseline during the maintenance phase was sustained by continuing ustekinumab administration from week 44 to week 92. The majority of subjects achieved a Mayo rectal bleeding subscore of 0, a Mayo bowel movement frequency subscore of 0 or 1, and an absolute stool count of 3 or less. - The decreases in CRP, fecal lactoferrin, and fecal calprotectin observed at baseline during the maintenance phase persisted from week 44 to week 92. - Improvements in health-related quality of life (IBDQ and SF-36) observed at baseline during the maintenance phase persisted from week 44 to week 92. ●Among the randomized subjects who received LTE, those who underwent dose adjustment showed some degree of benefit from the dose adjustment.

[0257] Ustekinumab induction-delayed responders who received LTE ●The subjects maintained symptomatic remission and partial Mayo remission from week 44 to week 92, achieved corticosteroid-free remission at week 92, maintained a decrease in inflammatory biomarkers from week 44 to week 92, and maintained improvements in health-related quality of life from week 44 to week 92. ●The clinical benefits observed in these subjects were similar to those observed in randomized subjects treated with ustekinumab q8w in the LTE group.

[0258] Efficacy and pharmacokinetics ●Generally, a high proportion (≥80%) of the subjects achieved symptomatic remission and partial Mayo remission at each concentration quartile. Therefore, in this population, which is considered to have benefited from maintenance therapy, no clear exposure-efficacy relationship was observed between serum ustekinumab concentrations and these efficacy endpoints.

[0259] Efficacy and immunogenicity ● The remission rate at week 92 was similar for randomized subjects who were antibody-positive for ustekinumab and those who were antibody-negative.

[0260] Safety results Among all subjects who received LTE, the overall safety profile from week 44 to week 96 was generally consistent with the known safety profile of ustekinumab.

[0261] ●The number of subjects who reported adverse events (AEs) was approximately the same for those treated with ustekinumab compared to those treated with placebo. The highest incidence of AEs among those who reported AEs was in the system-organ classes (SOC) of infections, invasiveness, and gastrointestinal disorders. The incidence rates of AEs (exceptions) in the SOC of infections and invasiveness per 100 person-years were 43.29, 48.91, and 46.48 in the placebo, ustekinumab q12w, and ustekinumab q8w groups, respectively. Nasopharyngitis was the most frequently reported AE, with incidence rates of 14.93, 21.55, and 19.83 in the placebo, ustekinumab q12w, and ustekinumab q8w groups, respectively. The incidence rates of AEs (exceptions) in the SOC (State of Computation) for gastrointestinal disorders per 100 person-years were 55.23, 34.82, and 31.53 in the placebo, ustekinumab q12w, and ustekinumab q8w groups, respectively. Ulcerative colitis was the most frequently reported AE, with a higher incidence rate among subjects in the placebo group (35.08) compared to the ustekinumab q12w and q8w groups (14.09 and 15.60, respectively). ●The number of all subjects who discontinued the study drug due to one or more adverse events (AEs) per 100 person-years in the follow-up period was 7.46 in the placebo group, 4.97 in the ustekinumab q12w group, and 4.23 in the ustekinumab q8w group, respectively. Ulcerative colitis was the most frequently reported AE leading to discontinuation, reported at 7.46, 2.49, and 2.28 cases per 100 person-years in the follow-up period. ●One patient died. This patient received a single dose of ustekinumab after dose adjustment from placebo. The direct cause of death was cardiac arrest and was determined to be unrelated to ustekinumab administration. Prior to cardiac arrest, the patient, who had multiple comorbidities, reported cytomegalovirus colitis, exacerbation of UC, and developmental delay. ●In the follow-up study, the number of subjects with at least one SAE per 100 person-years was 10.45 in the placebo group and 6.30 in the ustekinumab 90 mg combination SC group, which was similar to the subjects in the ustekinumab q12w group (5.80 cases) and q8w group (6.50 cases). The highest incidence of SAE per 100 person-years was associated with ulcerative colitis, at 5.22 in the placebo group and 1.63 in the ustekinumab combination group. ●The total number of subjects with one or more infections per 100 person-years in the follow-up study was 45.53 in the placebo group and 49.73 in the ustekinumab combination group, which was similar to the subjects in the ustekinumab q12w group (50.57 cases) and q8w group (50.71 cases). The most frequently reported infections occurring under treatment were nasopharyngitis (14.18 and 19.15) and upper respiratory tract infections (5.22 and 6.54), respectively, in the placebo group and the ustekinumab combination group. - In follow-up studies, the incidence rate of one or more infections requiring oral or parenteral antibiotic therapy per 100 person-years was 18.66 in the placebo group and 24.98 in the ustekinumab combination group. The most frequently reported infections requiring treatment with oral or parenteral antibiotics were nasopharyngitis, bronchitis, sinusitis, and upper respiratory tract infections. ●Severe infections were rarely reported. The incidence rate of one or more severe infections per 100 person-years in the follow-up study was 2.24 in the placebo group and 2.33 in the ustekinumab combination group. No specific events were reported in multiple subjects. ● No cases of active TB have been reported among patients treated with ustekinumab. ●Two cases of opportunistic infection were confirmed. In the placebo group, one patient who received a single dose of ustekinumab 90 mg after dose adjustment was diagnosed with cytomegalovirus colitis. The patient subsequently died due to cardiac arrest. In the ustekinumab q8w group, one patient was diagnosed with Listeria monocytogenes infection. This patient was reported to have recovered, albeit with sequelae. ● The proportion of all subjects who experienced one or more injection site reactions to ustekinumab was 2.2% (n=10), while the proportion of subjects who reported injection site reactions to placebo was 0.9% (n=4). In this study, no relationship was found between the expression of antibodies against ustekinumab and injection site reactions. ● No cases of anaphylactic or delayed-type hypersensitivity reactions were identified among patients treated with ustekinumab. ●In follow-up studies, the number of malignant tumors that developed per 100 person-years under treatment was 1.49 in the placebo group (one patient each: lentigo melanoma and basal cell carcinoma [BCC]) and 0.93 in the ustekinumab combination group. The rates were similar between the subjects in q12w (0.83; 1 patient, BCC development) and q8w (0.98; 2 patients, BCC development). One additional patient (ustekinumab q8w group) entered the LTE group but did not receive treatment because they were diagnosed with melanoma. ●Among all patients treated, three serious major adverse cardiovascular events (MACE; one fatal case) were reported (two patients in the randomized placebo group who received IV ustekinumab during the induction phase and had their dose adjusted to ustekinumab during the LTE period, and one patient in the delayed response group to ustekinumab induction [ustekinumab q8w administration]). Each patient had a confounding comorbidity at the time of the event. ● There was no significant difference in the proportion of subjects with maximum toxicity grade chemical and hematological laboratory values ​​of ≥1 after baseline between the placebo group and the respective ustekinumab groups. Grade 3 and grade 4 chemical and hematological laboratory values ​​were infrequent.

[0262] Overall, the safety profile of the randomized population was consistent with that observed in the entire population. In the limited number of populations that underwent dose adjustment to ustekinumab q8w, the safety profile was generally consistent with that observed in the population randomized during the maintenance phase of ustekinumab q8w. Serious adverse events and AEs leading to discontinuation of the study drug were rare in the population that underwent dose adjustment to ustekinumab q8w, with ulcerative colitis being the most frequently reported event.

[0263] The overall safety profile of ustekinumab in delayed-response subjects who received LTE was consistent with that observed in the randomized ustekinumab q8w group.

[0264] Results of healthcare economics and utilization of medical resources Among the randomized subjects who received LTE: ●The proportion of patients who underwent hospitalization or surgery related to UC disease from week 0 to week 96 of the induction period was low in both the placebo group (4.3% [5 patients]; patients in this group received a single IV dose of ustekinumab during the induction period) and the ustekinumab combination group (3.9% [11 patients]). ●At week 92, the improvement in productivity VAS scores observed at baseline during the maintenance phase was maintained in the ustekinumab group. ●At week 92, the mean percentage of WPAI-GH was maintained from the maintenance baseline in all four WPAI domains in the ustekinumab q12w and q8w groups, and additional improvements (i.e., reductions) were observed in the health-related work-related impairment rate, the health-related overall work-related impairment rate, and the health-related activity impairment rate in subjects in both ustekinumab groups.

[0265] Exam restrictions ● Participants in the LTE trial were selected because the principal investigator determined that continued administration could potentially provide benefits. This criterion may limit the generalizability of the trial results, as it only applies to patients who responded to and tolerated ustekinumab in the first year of treatment. ●To mimic real-world practices, the target group can change their concomitant medications at any time during the LTE period. ● Since the subjects who entered the LTE trial with placebo represented either long-term responders to ustekinumab induction or true placebo responders, a direct comparison of efficacy between the placebo group and the ustekinumab group was not performed. Furthermore, because the placebo subjects were discontinued from the trial when blinding was deblinded, the importance of a direct comparison between the placebo group and the ustekinumab group was limited. As a result, the reported clinical outcomes focused on efficacy indicators in the ustekinumab-treated subjects. ●Dose adjustments were made based on the principal investigator's clinical judgment regarding the disease activity of the subjects. Criteria specified in the study protocol (e.g., clinical flare based on partial Mayo scores) were not applied, and the interpretability of these data was limited because some subjects were in remission at the time of dose adjustment. ●Clinical outcomes in subpopulations based on the status of biological agent refractory (i.e., biological agent untreated, biological agent effective, and biological agent refractory) are presented to assess the consistency between outcomes in these subpopulations and outcomes in the overall population. However, because these analyses have limited sample sizes, these results must also be interpreted with caution.

[0266] conclusion ● Administration of ustekinumab 90 mg SC q12w and q8w maintained symptomatic remission or partial Mayo remission, as measured, up to the second year of administration. ●Even in the second year of administration, the effectiveness was maintained by a sustained decrease in disease-related inflammatory markers and a sustained improvement in health-related quality of life. ● No new safety signals were observed during the second year of maintenance therapy. - The safety profile is consistent with previously reported safety data up to the first year of administration in UC and with the overall safety profile of ustekinumab.

[0267] List of Abbreviations and Definitions of Terms

[0268] [Table 5]

[0269] 1. Introduction The Phase 3 development program for ustekinumab in the treatment of ulcerative colitis (UC) consists of two separate studies, an induction study and a maintenance study, conducted under the same clinical trial protocol (CNTO1275UCO3001). Both studies are Phase 3 randomized, double-blind, placebo-controlled, parallel-group, multicenter studies of ustekinumab in patients aged 18 years or older with moderate to severe active UC.

[0270] Introductory trials are conducted for subjects who have not achieved an adequate response or tolerated conventional or biological therapies.

[0271] Participants who demonstrated a clinical response to ustekinumab IV at week 8 or 16 of the induction trial were eligible to participate in a randomized discontinuation maintenance trial to evaluate the safety and efficacy of maintenance ustekinumab SC up to week 44. (Scope of the 96-week summary report)

[0272] This 96-week clinical trial summary of CNTO1275UCO3001 provides an overview of the efficacy, safety, pharmacokinetic (PK), and immunogenicity results from weeks 44 to 96 for subjects who continued long-term extension (LTE) administration in the maintenance study.

[0273] 2.Purpose The objective of the LTE trial was to evaluate the efficacy, safety, pharmacokinetics, and immunogenicity of ustekinumab administered for an additional three years to patients with moderate to severe active UC who had completed a 44-week maintenance study, and to patients whom the principal investigator determined would benefit from continued administration.

[0274] 3. Method 3.1 Overview of Phase 3 Program Design The induction and maintenance studies were phase 3 randomized, double-blind, placebo-controlled, parallel-group, multicenter trials of ustekinumab conducted under a single study protocol in patients aged 18 years or older with moderate to severe active ulcerative colitis (UC). The induction study included patients who had not achieved an adequate response or had poor tolerance to conventional or biological therapies (i.e., tumor necrosis factor [TNF] antagonists and integrin antagonists, vedolizumab). The maintenance study was a randomized discontinuation study for patients who had shown a clinical response to induction administration of ustekinumab IV. After the completion of the maintenance study (i.e., up to week 44), eligible patients received LTE for an additional three years under the same study protocol. Figure 1 shows a diagram of the study design.

[0275] Maintenance test design In the maintenance study, all enrolled subjects were respondents to the investigational drug administered in the induction study. The schema for the maintenance study is shown in Figure 2.

[0276] The primary population of the maintenance study consisted of subjects who demonstrated a clinical response to IV ustekinumab after induction. This population included: ● Patients who were randomized to ustekinumab (i.e., 130 mg IV or approximately 6 mg / kg IV) at week 0 of the induction trial and showed a clinical response at week 8 of the induction trial. ●A patient who was randomized to placebo at week 0 of the induction trial and did not show a clinical response at week 8 of the induction phase, but showed a clinical response at week 16 of the induction phase after receiving IV ustekinumab (approximately 6 mg / kg) at week 8 of the induction phase (placebo → ustekinumab approximately 6 mg / kg IV). Patients who demonstrated a clinical response to IV ustekinumab were randomized in a 1:1:1 ratio to one of three treatment groups (Table 6) at week 0 / baseline visit of the maintenance trial: ● Placebo SC ● Ustekinumab 90mg SC every 12 weeks (q12w) ● Ustekinumab 90mg SC every 8 weeks (q8w).

[0277] Eligible participants were assigned to a treatment group using a substitution block randomization method, stratified by clinical remission status at maintenance baseline (defined as Mayo score ≤2 and no individual subscores greater than 1) (yes / no), oral corticosteroid use at maintenance baseline (yes / no), and induction dose (placebo IV [induction phase 0 weeks] → ustekinumab approx. 6 mg / kg IV [induction phase 8 weeks], ustekinumab 130 mg IV [induction phase 0 weeks], or ustekinumab approx. 6 mg / kg IV [induction phase 0 weeks]).

[0278] The additional subjects entering the maintenance trial include the following. These subjects are not randomized and are not part of the primary population. ● Subjects who showed a clinical response to placebo IV induction and were administered placebo via SC (i.e., the placebo-induction responder group). ● Patients who were delayed responders to ustekinumab induction (i.e., those who did not show a clinical response to ustekinumab at week 8 of the induction period, but showed a clinical response at week 16 of the induction period after receiving ustekinumab via SC at week 8 of the induction period), and who were administered ustekinumab 90 mg SC q8w (i.e., the ustekinumab delayed responder group).

[0279] All subjects received their assigned dose of the SC study drug at their visit during week 0 of the maintenance phase. The maintenance study continued until week 44.

[0280] Design of long-term continuous administration studies After the safety and efficacy evaluation at week 44 was completed, patients who the principal investigator determined would benefit from continued administration were given the opportunity to participate in the Long-Term Recovery (LTE). The LTE was initiated after the evaluation recorded at the 44-week maintenance visit (M-44) and continued until week 220.

[0281] During the LTE period, participants continued the same dosing regimen as at the end of the maintenance study (placebo, ustekinumab 90 mg SC q12w, or ustekinumab 90 mg SC q8w), with the first dose in the LTE administered at week 48.

[0282] During the LTE period, all subjects needed to be evaluated for worsening UC disease activity based on the clinical judgment of the principal investigator. Among the primary population (i.e., subjects randomized at week 0 of the maintenance phase), those who experienced worsening UC disease activity received a single dose adjustment as follows: ● Placebo SC → Ustekinumab 90mg SC q8w ● Ustekinumab 90mg SC q12w → Ustekinumab 90mg SC q8w ● Ustekinumab 90mg SC q8w → Ustekinumab 90mg SC q8w continued

[0283] The patient's first visit to the hospital to consider dose adjustment was at week 56. The patient was able to undergo one dose adjustment during the LTE period.

[0284] The Interaction Web Response System (IWRS) confirmed that the frequency of ustekinumab SC administration did not exceed q8w. For example, among subjects randomized to the ustekinumab 90mg SC q12w group, those whose disease activity was judged to have worsened by the principal investigator received ustekinumab 90mg SC at the current visit only if the last dose of ustekinumab had been administered at least 8 weeks prior to the current visit. If the last dose of ustekinumab 90mg SC was less than 8 weeks prior, the next dose of ustekinumab 90mg SC was to be initiated at the next scheduled visit at least 8 weeks after the previous dose of ustekinumab. From week 56 of administration, the principal investigator was instructed to assess the possibility of worsening UC disease activity in the subject according to the study protocol, and if the subject's dose had not been adjusted, to assess the need for dose adjustment based on the principal investigator's clinical judgment. At each facility, participants entered "yes" or "no" in response to the question of whether the subject needed dose adjustment. If the answer was "yes," dose adjustments and drug distribution were managed in a blinded manner using the IWRS until the study was deblinded. After the study was deblinded, subjects who had received ustekinumab q12w could receive dose adjustments to ustekinumab q8w if their dose adjustments had not yet been performed.

[0285] Dose adjustments during the LTE period were not applied to subjects not included in the primary population (placebo-initiated responders, ustekinumab delayed responders).

[0286] For patients in whom the principal investigator determined that no improvement in UC disease activity was observed by 16 weeks after dose adjustment, the administration of the study drug was discontinued.

[0287] During the LTE period, all concomitant medications, including those specific to UC (with the exception of certain prohibited medications listed below), were permitted at the discretion of the principal investigator.

[0288] Efficacy assessments during the LTE period included partial Mayo scores, inflammatory markers, and corticosteroid use. Full Mayo scores (including endoscopy) were assessed at the final efficacy visit at week 200, at discontinuation of the study drug, or at the end of study participation. Selected patient-reported outcomes and health economic data were also collected. Safety assessments included evaluation of adverse events (AEs) and periodic laboratory analyses. The final safety assessment was conducted at the visit at week 220 or approximately 20 weeks after the last dose of the study drug (for subjects who did not discontinue study participation). All study assessments conducted during the LTE period are documented in the Time and Events Schedule of the study protocol.

[0289] During the LTE period, the blinded trial was maintained until the last participant in the maintenance trial completed their visit evaluation at week 44 (M-44) and the week 44 analysis was completed. Therefore, participants continued to receive the study drug at their monthly visits until then. After the trial was unblinded at the clinical trial sites, participants who received placebo discontinued participation in the trial, and participants who received ustekinumab continued to receive ustekinumab, but their trial visits were scheduled according to their administration regimen (either q8w or q12w, if appropriate for their regimen).

[0290] The sponsor blinded dosing assignments in the maintenance study until a disease-blocking event (DBL) occurred at week 44. To minimize bias and protect the integrity of the clinical program, blinding of dosing assignments was maintained for the sites, site monitors, and participants in this study (both introductory and maintenance studies) until the analysis at week 44 was completed. Participants entered the LTE with their assigned maintenance dosing regimen (e.g., q8w or q12w), receiving injections of the study drug every four weeks (except week 52), with the first injection administered at week 48, and blinding was maintained. After the study blinding was lifted for the sites, participants who received placebo discontinued participation in the study, and participants who received ustekinumab continued to receive ustekinumab, but their visits were scheduled according to their dosing regimen (q8w or q12w until week 200, as appropriate).

[0291] During the LTE period, all concomitant medications, including those specific to UC (excluding the prohibited medications listed below), were permitted at the discretion of the principal investigator. Furthermore, prohibited therapies were not to be used as rescue medications.

[0292] Test evaluation The following tests and evaluations were conducted: Pharmacokinetics: Serum ustekinumab concentration Immunogenicity: Antibodies against ustekinumab Effectiveness: Partial Mayo score, C-reactive protein (CRP), fecal lactoferrin, fecal calprotectin, and use of corticosteroids. Health-related quality of life: Inflammatory Bowel Disease Questionnaire (IBDQ), 36-item Short-Form Health Survey (SF-36) Health Economics: Hospitalizations and surgeries related to UC disease; Productivity Visual Analog Scale (VAS), and Work Productivity and Activity Impairment Questionnaire - General Health (WPAI-GH) Safety: AEs, serious adverse events (SAEs), infections, injection site reactions, allergic reactions, hematological and chemical parameters, vital signs, physical examination, and early detection of tuberculosis (TB).

[0293] Pharmacokinetics and immunogenicity Blood samples for measuring serum ustekinumab concentrations and ustekinumab immunogenicity (antibodies against ustekinumab) were collected from all subjects as indicated in the LTE time and event schedule. The analysis was performed as previously presented in UCO3001 44W.

[0294] Effectiveness evaluation Efficacy evaluations up to week 92 for those entering the LTE were conducted as outlined in the LTE time and event schedule, and included partial Mayo scores, CRP, fecal lactoferrin and fecal calprotectin, corticosteroid use, IBDQ, and SF-36.

[0295] Descriptions of individual efficacy evaluations were previously presented in UCO3001 44W.

[0296] Criteria for effectiveness The efficacy endpoint was defined as follows: ● Clinical remission: Mayo score of 2 or less, and no individual subscores exceeding 1. ●Symptomatic remission: Mayo bowel movement frequency subscore is 0 or 1, rectal bleeding subscore is 0 ●Partial Mayo remission: Partial Mayo score of 2 or less ● IBDQ remission: IBDQ level of 170 or higher ● Normalization of CRP concentration: CRP concentration is 3 mg / L or less. ● Normalization of fecal lactoferrin concentration: Fecal lactoferrin concentration should be 7.24 μg / g or less. ● Normalization of fecal calprotectin concentration: Fecal calprotectin concentration should be 250 mg / kg or less.

[0297] Safety evaluation Safety up to week 96 was assessed based on adverse events (AEs) and laboratory results (i.e., hematology and serology), as previously described in UCO3001 44W. Except for laboratory data, data on safety variables were recorded or attached to electronic case report forms. Laboratory data were collected and stored in electronic file format. The timing of all safety procedures was documented in the LTE time and event schedule in the study protocol.

[0298] Safety data obtained during the trial were regularly reviewed by an open-label, independent DMC until week 44 of the DBL (Diagnosis-Based Learning).

[0299] Health economics and utilization of medical resources Healthcare resource utilization data, including hospitalizations and surgeries related to UC, was collected. Furthermore, the potential impact of ustekinumab on work limitations and daily productivity in the target population was assessed using WPAI-GH and productivity VAS, respectively.

[0300] Data Quality Assurance The trial was monitored in accordance with the sponsor's current standard operating procedures for clinical trial monitoring.

[0301] statistical methods The primary objective of the LTE trial was to ensure that subjects reaching week 44 of the maintenance trial could continue receiving the investigational drug without interruption. The main objectives of this trial report are to evaluate efficacy from the end of the maintenance trial (week 44) to week 92 of the LTE (the final efficacy assessment before week 96), and safety from the end of the maintenance trial to week 96 of the LTE. However, data prior to week 44 is also included.

[0302] It is important to note that the subjects entered the long term (LTE) based on the investigator's judgment that continued administration would provide a benefit. Furthermore, the placebo group represents a subgroup of UC patients who were either long-term responders to ustekinumab induction therapy (i.e., re-randomized during the placebo maintenance phase) or placebo induction responders with a long disease latency period. For these reasons, and because the placebo subjects were to discontinue participation in the trial after deblinding, a direct comparison of the results between the placebo group and the ustekinumab group was considered confounding. Therefore, no statistical comparison was performed.

[0303] Continuous variables were summarized using descriptive statistics (e.g., mean, median, standard deviation, interquartile range, minimum, and maximum). Categorical variables were summarized using counts and proportions.

[0304] Planned analysis The planned analysis for LTE is as follows:

[0305] Population to be analyzed Effectiveness Summary of efficacy was provided for randomized subjects who received LTE at week 0 of the maintenance trial. Summary of selected efficacy was also provided for subjects who underwent dose adjustments during the LTE period, and for non-randomized subjects who received LTE at week 0 of the maintenance trial. Furthermore, regardless of whether subjects received LTE, separate summary of efficacy for symptomatic remission and partial Mayo remission endpoints was provided for all randomized and non-randomized subjects at baseline during the maintenance phase.

[0306] The primary population for the efficacy summary consisted of randomized subjects who received LTE.

[0307] safety The safety summary was based on all subjects who received at least one dose of the study drug in the LTE. Additional summaries were also provided based on the randomization status up to dose adjustment (randomized or non-randomized in the maintenance study) and the randomized subjects, including data after dose adjustment.

[0308] The target population for administration in LTE was the main population for the safety summary.

[0309] Pharmacokinetics Pharmacokinetic analyses were performed on all subjects (including both randomized and non-randomized subjects) who received at least one dose of ustekinumab during the LTE period. Analysis was also conducted on subjects who underwent dose adjustments during the LTE period.

[0310] immunogenicity Immunogenicity analysis was performed on all subjects who received LTE, received at least one ustekinumab dose, and had at least one sample taken after the first ustekinumab dose for antibody detection against ustekinumab.

[0311] Pharmacokinetics Serum concentrations at weeks 44, 68, and 92 were aggregated for each administration.

[0312] In the list containing concentration data, all concentrations below the minimum quantifiable concentration were displayed as such. Concentrations below the minimum quantifiable concentration were treated as zero in the summary statistics.

[0313] In the summary of serum ustekinumab concentrations including data up to week 44, the following data from the time of onset to week 44 were excluded: (1) discontinuation of the study drug, (2) skipping an injection, (3) receiving an incomplete injection, (4) receiving an incorrect injection, (5) receiving an additional injection, and / or (6) receiving commercially available ustekinumab. Furthermore, PK samples collected outside the scheduled visit period (±10 days of each scheduled visit) were excluded from the summary. These exclusion rules did not apply to data after week 44.

[0314] immunogenicity The expression rate of antibodies against ustekinumab was aggregated for subjects who received administration up to week 96 in the LTE and had a suitable sample for detecting antibodies against ustekinumab (i.e., subjects from whom at least one sample was obtained after the first dose of ustekinumab).

[0315] Based on randomized maintenance-phase subjects who received ustekinumab via LTE, serum ustekinumab concentrations based on antibody status against ustekinumab at weeks 44, 68, and 92 were aggregated by treatment group up to week 96.

[0316] A list of subjects who tested positive for antibodies to ustekinumab from week 0 to week 96 of the induction period was provided.

[0317] Effectiveness Data Processing Rules Rules for Ineffective Administration: Unless otherwise specified, subjects who underwent ostomy or colectomy, lacked efficacy, discontinued the study drug due to adverse events (such as UC exacerbation), or underwent dose adjustments before designated hospital visits (occurring only after week 56) were considered to have had an ineffective administration from the time the event occurred.

[0318] For binary endpoints, patients who did not respond to treatment were considered to have failed to achieve each endpoint from the time of treatment failure onward. For continuous endpoints, patients who did not respond to treatment were given baseline values ​​from the time of treatment failure.

[0319] Missing Value Rules: For objects with missing values, unless otherwise specified, the last observed value was carried over for continuous endpoints, except for the last available Mayo subscore which was carried over as a partial Mayo score. For binary endpoints, objects with missing values ​​were considered not to have achieved each endpoint.

[0320] The rule for administration failure takes precedence over the rule for missing values. In other words, if an event of administration failure occurs in a subject, the baseline value for the continuous endpoint is assigned from the time of administration failure, and the subject is considered not to have achieved each of the binary endpoints, regardless of observed or missing values.

[0321] analysis method In this CSR, the following three analytical methods were employed for subjects who received LTE: ● As-observed analysis: Data up to week 92 or until dose adjustment was aggregated, the ineffectiveness rule was applied, and subjects with missing data that did not result in ineffectiveness before the specified analysis time were excluded. ●Intention to Treat (ITT): Data up to week 92 was aggregated, and rules for ineffective administration and missing values ​​were applied. ●Dose adjustment as a treatment strategy: Except for temporarily suspending the criteria for ineffectiveness due to dose adjustment, this is the same as the corresponding ITT analysis method (i.e., subjects who underwent dose adjustment are not considered to have received ineffective treatment). The other analysis rules were the same.

[0322] In the as-observed analysis method, only subjects for whom data was available at each analysis time point, or subjects who had previously been ineffective (considered non-responders), were included in the analysis. This approach was considered reasonable because it excluded only patients with missing data unrelated to treatment ineffectiveness (likely randomly missing).

[0323] In the ITT analysis method, the number of subjects included in the analysis was fixed over time. It was expected that the proportion of subjects achieving the binary endpoint would decrease over time, as the number of subjects undergoing dose adjustments (based on criteria for ineffectiveness) or discontinuing the study drug (whether due to lack of efficacy or worsening UC) was anticipated. Therefore, the ITT analysis method was considered conservative.

[0324] For efficacy analyses, conservative ITT analysis was used by default. However, for key efficacy endpoints such as symptomatic remission, partial Mayo remission, and change from baseline in partial Mayo scores, analyses based on as-observed analysis were performed and were considered to more reasonably reflect efficacy in LTE.

[0325] The analysis method based on "dose adjustment as a treatment strategy" was considered practical because it reflects clinical practice where administration is optimized by increasing the dose or frequency of administration.

[0326] Clinical endpoints Table 5 lists the clinical endpoints aggregated in this CSR, along with the relevant analysis populations and methods based on the subjects who received LTE. Clinical remission, as referenced in the analysis, was based on the global definition (Mayo score of 2 or less, with no individual subscores exceeding 1).

[0327] In addition to summaries based on subjects who received LTE, symptomatic remission and partial Mayo remission were separately aggregated for all randomized and non-randomized subjects at baseline during the maintenance phase, regardless of whether they received LTE, using an analysis method based on "dose adjustment as a treatment strategy" (randomized subjects only) and an ITT analysis method. In this type of analysis, in accordance with the rules for ineffectiveness applied in UCO3001 W44 CSR, subjects who were prohibited from changing their UC medication, who underwent ostomy or colectomy, who used rescue medication after a clinical flare, or who discontinued the study drug before the 44-week visit due to lack of efficacy or UC exacerbation as an AE were considered to have had an ineffective episode before or at 44 weeks. Subjects who underwent ostomy or colectomy after 44 weeks of treatment, or who discontinued the study drug due to lack of efficacy or UC exacerbation as an AE (or who underwent dose adjustment using the ITT analysis method) were considered to have had an ineffective episode from the time of the event.

[0328] [Table 6]

[0329] Inflammatory biomarkers Based on the ITT analysis method, the following endpoints were aggregated for both randomized and non-randomized subjects at week 0 of the maintenance trial who received LTE. ● Changes in CRP, fecal lactoferrin, and fecal calprotectin concentrations from baseline (maintenance and induction phases) over the period up to week 92. ● In subjects with abnormal CRP, fecal lactoferrin, and fecal calprotectin levels at baseline during the induction phase, normalization of CRP, fecal lactoferrin, and fecal calprotectin levels was achieved up to week 92.

[0330] Health-related quality of life Based on the ITT analysis method, the following endpoints were aggregated from randomized subjects at week 0 of the maintenance trial who received LTE. ● IBDQ - Changes in IBDQ score and each of the four IBDQ items from baseline (maintenance and induction phases) up to week 92. - Improvement of 16 points or more from the IBDQ baseline during the period up to week 92. - Among subjects whose IBDQ score improved by 16 points or more from the introduction baseline at the maintenance baseline, the improvement of 16 points or more from the introduction baseline over time up to week 92. - Among subjects whose IBDQ score improved by 16 points or more at the maintenance baseline (from the introduction baseline), the following improvement of 16 points or more from the introduction baseline was observed at both week 44 and week 92. - Of the subjects whose IBDQ improved by 16 points or more (from baseline) at week 44, those whose IBDQ improved by 16 points or more from baseline at both week 68 and week 92 - IBDQ remission over a period of up to week 92 - Among subjects who achieved IBDQ remission at baseline during the maintenance phase, IBDQ remission for the period up to week 92 - Among subjects who achieved IBDQ remission at baseline during the maintenance phase, the time-series IBDQ remission at both weeks 44 and 92 was measured. - IBDQ remission over time at both weeks 68 and 92 in subjects who achieved IBDQ remission at week 44. ●SF-36 - Changes from baseline (maintenance and induction phases) in SF-36 Physical Component Summary (PCS) and Mental Component Summary (MCS) scores over the period up to week 92. - Over the period up to week 92, patients showed an improvement of 5 points or more from baseline in both the SF-36 PCS and SF-36 MCS. - Among subjects whose SF-36PCS and SF-36 MCS improved by 5 points or more at the maintenance baseline, those who showed an improvement of 5 points or more from the induction baseline in both SF-36PCS and SF-36 MCS over the period up to week 92. - Among subjects whose SF-36PCS and MCS improved by 5 points or more at the maintenance baseline, those who showed 5 points or more improvement from the induction baseline in both SF-36PCS and SF-36 MCS at both weeks 44 and 92. -Among subjects whose SF-36PCS and MCS improved by 5 points or more at week 44, those who showed an improvement of 5 points or more from baseline in both SF-36PCS and SF-36 MCS at both weeks 68 and 92.

[0331] Furthermore, among the non-randomized subjects at week 0 of the maintenance trial, the following endpoints were aggregated for those who received LTE based on the ITT analysis method: - Change from maintenance baseline in IBDQ score over the period up to week 92 - Improvement of 16 points or more from the IBDQ baseline over a period of up to week 92. - IBDQ remission for up to week 92 - Changes from maintenance baseline in SF-36PCS and MCS over the period up to week 92 - Over the period up to week 92, patients showed improvement of 5 points or more from baseline in both the SF-36 PCS and SF-36 MCS.

[0332] Patients who underwent dose adjustments during long-term continuous administration For randomized subjects who underwent dose adjustments before or at week 76 and for whom data were available for at least 16 weeks after dose adjustment, data from the time of dose adjustment and at least 16 weeks after the initial visit were aggregated for the following endpoints: ●Symptomatic remission ● Partial Mayo remission ● Partial Mayo score ●CRP (mg / L) ● Calprotectin in feces (mg / kg) ●Lactoferrin in feces (μg / g)

[0333] Furthermore, a similar overview was provided for symptomatic remission and partial Mayo remission based on the biological agent refusal profile (biological agent untreated, biological agent effective, and biological agent refusal). In addition, subjects who were not in symptomatic remission / partial Mayo remission at the time of dose adjustment but were in symptomatic remission / partial Mayo remission at their first visit at least 16 weeks after dose adjustment were also aggregated. In all of these analyses, subjects who underwent ostomy or colectomy, or who discontinued the study drug due to lack of efficacy or worsening of UC as an adverse event, were considered refusal of treatment from the time of the event onward.

[0334] Efficacy and pharmacokinetics The following efficacy endpoints were aggregated based on ustekinumab concentrations at week 92 (≥1st quartile, ≥1st quartile and <2nd quartile, ≥2nd quartile and <3rd quartile, ≥3rd quartile) and the mean trough serum ustekinumab concentration up to week 96, based on randomized maintenance patients who received ustekinumab in the LTE and did not undergo dose adjustment. ●Symptomatic remission at 92 weeks ● Partial Mayo remission at 92 weeks ● Changes from maintenance baseline in CRP, fecal lactoferrin, and fecal calprotectin levels at week 92 ● Normalization of CRP, fecal lactoferrin, and fecal calprotectin at week 92 in subjects who had abnormal CRP, fecal lactoferrin, and fecal calprotectin levels, respectively, at baseline during the induction phase.

[0335] Efficacy and immunogenicity In randomized patients receiving LTE during the maintenance phase, the relationship between antibody status against ustekinumab up to week 96 and partial Mayo remission and symptomatic remission at week 92 was examined.

[0336] safety This CSR focused the safety summary on all subjects who received at least one dose of the study drug in the LTE. The safety summary was primarily based on data from weeks 44 to 96, although some key safety analyses included data prior to week 44. Additional summaries were also provided based on the randomization status up to dose adjustment (randomized or non-randomized in the maintenance study) and the randomized subjects, including data after dose adjustment.

[0337] Adverse events AEs that occurred under administration were coded according to the Medical Regulatory Terminology (MedDRA) version 21.1, using the term closest to the principal investigator's terminology as the lower-level term (LLT), closely related LLT groups as preferred terms (PT), and the broad categories including related PTs as the organ-specific broad categories (SOC).

[0338] The proportion of subjects who experienced one or more adverse events (AEs) under the following treatment regimens was aggregated by treatment group: ● Any AE ●SAE ● AEs that led to discontinuation of the investigational drug ● Injection site reaction ● Infectious diseases and serious infectious diseases

[0339] Injection site reactions are any adverse reactions at the injection site of the SC test drug, as recorded by the principal investigator in the electronic case report form (eCRF) as AEs (and injection site reactions).

[0340] Infections were defined as any AE that the principal investigator identified as an infection on the eCRF.

[0341] The list includes SAEs that occurred during administration, AEs that led to discontinuation of the study drug, malignancies, major adverse cardiovascular events (MACEs), embolic and thrombotic events, and death.

[0342] Furthermore, the number of events per 100 person-years and the number of individuals with events per 100 person-years in the follow-up study were aggregated to adjust for potential differences during the follow-up period.

[0343] Furthermore, this CSR (Clinical Survey) includes the incidence rate of malignant tumors.

[0344] Clinical Tests The maximum values ​​of the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE) toxicity grades after baseline from week 44 to week 96 were aggregated by clinical laboratory and treatment group.

[0345] In addition, clinical laboratory values ​​of CTCAE grade 2 or higher are listed. The NCI-CTCAE toxicity grades are based on NCI-CTCAE version 4.03.

[0346] Safety and immunogenicity Furthermore, for subjects who received ustekinumab SC during the LTE period, the relationship between injection site reactions from week 44 to week 96 and antibody status against ustekinumab up to week 96 was examined.

[0347] Utilization of medical resources and health economics Based on randomized subjects at week 0 of the maintenance trial who received LTE, the following endpoint summaries were provided. No data processing rules were applied. ●Percentage of subjects who underwent hospitalization, surgery, or both related to UC disease between week 0 and week 96 of the introductory period. ●Percentage of subjects who underwent hospitalization, surgery, or both related to UC disease between week 0 and week 96 of the maintenance phase. ●Percentage of subjects who underwent hospitalization, surgery, or both related to UC between weeks 44 and 96. ● Changes in productivity VAS from baseline (maintenance and induction phases) over the period up to week 92. ● Changes from baseline (maintenance and induction phases) in the prevalence of each of the four disability categories due to WPAI-GH over the period up to week 92.

[0348] Target and treatment information Target deployment and test completion / departure information Distribution of registered subjects by treatment group and region The patient placement for the first 44 weeks of this study is described in UCO3001 44W CSR. Safety and efficacy evaluations were completed at 44 weeks, and a total of 588 patients received LTE if the principal investigator determined that continued administration would be beneficial. Of these, 399 patients were in the primary maintenance population (i.e., those who showed a clinical response to IV ustekinumab induction and were randomized at week 0 of the maintenance phase). Figure 3 and 189 patients were not included in the primary maintenance population (i.e., placebo induction responders and ustekinumab induction delayed responders [non-randomized subjects]; Figure 3).

[0349] Of the subjects randomized at the maintenance baseline, the 399 who received treatment during the LTE period were as follows (Figure 3): ● Placebo SC: 115 cases ● Ustekinumab 90mg SC q12w: 141 cases ● Ustekinumab 90mg SC q8w: 143 cases

[0350] Among the primary population of the maintenance study (i.e., those randomized at week 0), those who received LTE and experienced worsening UC disease activity during the LTE period received a single dose adjustment to ustekinumab 90 mg q8w (Section 0). A total of 32.6% (130 patients) of the randomized population underwent a dose adjustment during the LTE period.

[0351] The 189 non-randomized patients who received treatment during the maintenance phase in the LTE period are as follows (Figure 3): ● Placebo SC: 73 subjects who showed a clinical response to placebo IV induction (placebo IV responseers) continued to receive placebo SC during the maintenance period and LTE period until blinding of the study was lifted and the study was discontinued (see Section 0 for details). ●Ustekinumab 90 mg q8w: 116 patients with delayed response to ustekinumab induction (i.e., those who did not show a clinical response to ustekinumab at week 8 of the induction phase, but showed a clinical response at week 16 of the induction phase after receiving ustekinumab via SC at week 8 of the induction phase) were continuously administered ustekinumab 90 mg SC q8w during the maintenance period and until LTE.

[0352] The 588 participants, including all randomized and non-randomized subjects, came from 196 different locations: Asia (14.8%), Eastern Europe (44.9%), and other regions (including North America, Western Europe, Israel, Australia, and New Zealand) (40.3%).

[0353] Trial participation status up to week 96 Discontinuation of the test agent Figure 4 shows the number of subjects who received LTE and discontinued the study drug by week 96. For subjects who discontinued the study drug, a safety follow-up was conducted for approximately 20 weeks from the last dose of the study drug.

[0354] Randomized subjects Among the randomized population, 17.8% (71 patients; Figure 4) discontinued the study drug by week 96.

[0355] The proportion of patients who discontinued the study drug in each treatment group was 40.9% in the placebo group (including patients who discontinued after deblinding of the study [29.6%]) and 8.5% in the ustekinumab combination group. The proportions were similar in the ustekinumab q12w and q8w groups, respectively (9.2% and 7.7%, respectively). The most common reasons for discontinuing the study drug in the ustekinumab combination group were adverse events due to worsening of UC (2.5% [7 cases]) and other reasons (2.5% [7 cases]; most of which were reported as withdrawal of consent).

[0356] Non-randomized subjects In the non-randomized group, 27.5% (52 patients; Figure 4) discontinued the study drug by week 96.

[0357] The proportion of non-randomized subjects who discontinued the study drug from each treatment group was 64.4% (47 subjects) in the placebo-initiated responders group (including subjects who discontinued after deblinding of the study [39.7% (29 subjects)]) and 4.3% (5 subjects) in the ustekinumab delayed-initiated responders group (Table 6). The most common reason for discontinuing the study drug in the ustekinumab delayed-initiated responders group was "other" (1.7% [2 subjects]); both were reported as withdrawal of consent).

[0358] [Table 7]

[0359] End of participation in the exam The following is a summary of participants who took part in the LTE trial and discontinued their participation by week 96.

[0360] Randomized subjects By week 96, 11.4% (46 patients) of randomized subjects discontinued participation in the trial. This was 32.5% (38 patients) in the placebo group (25.6% [30 patients] discontinued after deblinding of the trial) and 2.8% (8 patients) in the ustekinumab combination group. The most common reason for discontinuing participation in the trial in the ustekinumab combination group was withdrawal of consent (2.4% [7 patients]).

[0361] Non-randomized subjects The proportion of non-randomized participants who discontinued the trial by week 96 was 47.9% (35 patients) in the placebo-initiated responders group (28.8% [21 patients] discontinued after deblinding of the trial) and 5.0% (6 patients) in the ustekinumab-initiated delayed responders group. The most common reason for discontinuing participation in the ustekinumab-initiated delayed responders group was withdrawal of consent (4.2% [5 patients]).

[0362] The results for subjects who received LTE and discontinued participation in the study by week 96 were similar to the results for subjects who received LTE, as described above.

[0363] Unblinding of the study drug from week 44 to week 96. Of the three patients who received LTE (all from the randomized population), two in the placebo group and one in the ustekinumab q8w group were unblinded before the deblinding of the trial from week 44 to week 96. All three patients were deblinded by their respective institutions for the purpose of further medical management after discontinuation of the study drug. All three patients completed the early termination visits required by the clinical trial protocol.

[0364] Demographic characteristics and baseline characteristics Demographic and baseline clinical disease characteristics were based on the population that received LTE. The primary analysis population was the randomized population; therefore, the data presentation focuses on these subjects. Data from non-randomized subjects are also provided, focusing on the ustekinumab delayed response group.

[0365] demographics Randomized subjects Of the randomized participants, 58.1% were male, 74.4% were white, the median age was 40.0 years, and the median weight was 71.60 kg.

[0366] Non-randomized subjects The demographic characteristics observed in the non-randomized population were largely similar to those observed in the randomized population. However, delayed responses to ustekinumab were more likely to be male compared to the randomized population.

[0367] Clinical disease characteristics The baseline disease characteristics during the induction and maintenance phases of randomized subjects receiving LTE were consistent with those of the entire randomized population in the maintenance trial.

[0368] Disease characteristics at 44 weeks of maintenance Randomized subjects The clinical disease characteristics at week 44 of randomized subjects receiving LTE were generally similar in the ustekinumab q12w and q8w groups, with the placebo group showing higher numerical values ​​(e.g., Mayo score, CRP concentration) or lower values ​​(e.g., those in remission), indicating greater disease activity in the placebo group. The data for the ustekinumab q12w and q8w groups are shown below, respectively. ●Percentage of subjects in clinical remission (world definition): 46.1% and 52.4% ●Percentage of patients who underwent endoscopic treatment: 56.7% and 61.5% ● Average Mayo score: 2.6 and 2.4 ●Median IBDQ scores: 193.0 and 194.0 ●Median CRP concentration: 1.47 mg / L and 1.41 mg / L ●Median fecal calprotectin concentration: 118.00 mg / kg and 158.00 mg / kg ●Median fecal lactoferrin concentration: 9.08 μg / g and 13.30 μg / g

[0369] The clinical disease characteristics at week 44 in the randomized population in the placebo group receiving LTE were as follows: ●Percentage of subjects in clinical remission (world definition): 34.8% ●Percentage of patients who underwent endoscopic treatment: 47.8% ●Average Mayo score: 3.2 ●Median IBDQ score: 185.0 ●Median CRP concentration: 2.56 mg / L ●Median fecal calprotectin concentration: 368.00 mg / kg ●Median lactoferrin concentration in feces: 28.95 μg / g

[0370] Non-randomized subjects The clinical disease characteristics at week 44 of the ustekinumab delayed response group (ustekinumab administered at q8w during the LTE period) indicated higher disease activity compared to the clinical disease characteristics of the randomized ustekinumab q8w group (e.g., fewer subjects achieving remission in the clinical efficacy endpoint, higher levels of inflammatory biomarkers). The data for each group are shown below: ●Percentage of subjects in clinical remission (world definition): 38.8% and 52.4% ●Percentage of patients who underwent endoscopic treatment: 47.4% and 61.5% ● Average Mayo scores: 3.2 and 2.4 ●Median IBDQ scores: 189.5 and 194.0 ●Median CRP concentration: 1.72 mg / L and 1.41 mg / L ●Median fecal calprotectin concentration: 324.00 mg / kg and 158.00 mg / kg ●Median fecal lactoferrin concentration: 30.06 μg / g and 13.30 μg / g

[0371] Prior treatment and combination therapy The presented concomitant UC medications and UC-related medication history are from week 0 of the introductory trial for all subjects who received LTE.

[0372] Combination therapy Randomized subjects At baseline during the induction phase, 90.5% of randomized subjects receiving LTE were also receiving concomitant UC medications. The overall proportions of subjects receiving corticosteroids, immunomodulators, and aminosalicylates were 50.1%, 29.3%, and 73.9%, respectively.

[0373] [Table 8]

[0374] The proportion of randomized subjects receiving corticosteroids and immunomodulatory agents at baseline during the induction phase was balanced among the ustekinumab treatment groups, but the proportion of aminosalicylic acid recipients was 74.8%, 81.6%, and 65.7% in the placebo, ustekinumab q12w, and ustekinumab q8w groups, respectively.

[0375] Non-randomized subjects The use of concomitant UC medications at baseline during the induction phase in the ustekinumab delayed response group was generally consistent with the use of concomitant UC medications in the randomized population from the ustekinumab q8w group.

[0376] Medical history Randomized subjects The majority (93.7%) of patients who received LTE and were randomized during the maintenance phase either had an inadequate response to or poor tolerance of corticosteroids and / or 6-mercaptopurine / azathioprine (6-MP / AZA) or exhibited corticosteroid dependence at baseline during the induction phase. A history of response to and intolerance to UC drugs was similar across all treatment groups. Of the randomized patients, 74.9% were refractory, dependent on, or intolerant to corticosteroids, and 54.9% were refractory or intolerant to 6-MP / AZA.

[0377] Among the randomized subjects receiving LTE in the maintenance phase, 55.9% had no history of biological agent failure at baseline during the induction phase (53.1% were biological treatment newcomers, and 2.8% had received biological treatment but had no record of biological agent failure). The proportion of randomized subjects with a recorded history of biological agent failure was lower in the ustekinumab q12w group (37.6%) compared to the ustekinumab q8w group (49.7%).

[0378] Overall, among the subjects randomized during the maintenance phase who received LTE, the following had a history of biological agent refractory: ●43.9% were ineffective against at least one type of biological agent for anti-TNF (regardless of vedolizumab). -32.1% were refractory to biological agents targeting only anti-TNF (not vedolizumab). ●12.0% were refractory to vedolizumab (regardless of anti-TNF). ●11.8% were unresponsive to any anti-TNF and vedolizumab biological agents.

[0379] Non-randomized subjects The response to and intolerance to UC medications, as well as the history of UC medication use, in the ustekinumab delayed response group who received treatment during the LTE period were generally consistent with that of the randomized population from the ustekinumab q8w group.

[0380] Randomized subjects in the maintenance phase after dose adjustment. During the LTE period, dose adjustments were applied to maintenance-phase randomized subjects who, based on the clinical judgment of the principal investigator, were judged to have experienced a worsening of UC disease activity as early as week 56.

[0381] Target distribution by treatment group Of the randomized population, a total of 32.6% (130 patients) underwent the following dose adjustments: ●Of those randomized to placebo, 46.1% (53 patients) received dose adjustments to the ustekinumab 90 mg SC q8w regimen. Of the patients randomized to ustekinumab 90 mg SC q12w, 28.4% (40 patients) had their dose adjusted to the ustekinumab 90 mg SC q8w regimen. ● Of the patients randomized to ustekinumab 90 mg SC q8w, 25.9% (37 patients) underwent sham dose adjustment (continuation of the same administration regimen).

[0382] The vast majority of patients who underwent dose adjustments had their doses adjusted by week 68.

[0383] Trial participation status up to week 96 Discontinuation of the test agent Of the 130 patients who underwent dose adjustments during the LTE period, 19 patients (14.6%) discontinued the study drug. The percentages of patients who discontinued the study drug were 15.1%, 17.5%, and 10.8% for the placebo group → ustekinumab q8w group, ustekinumab q12w group → ustekinumab q8w group, and ustekinumab q8w group → ustekinumab q8w group, respectively. The most common reason for discontinuing the study drug was adverse events due to worsening of ulcerative colitis (5 patients [9.4%], 5 patients [12.5%], and 0 patients for the placebo group → ustekinumab q8w group, ustekinumab q12w group → ustekinumab q8w group, and ustekinumab q8w group → ustekinumab q8w group, respectively).

[0384] End of participation in the exam Of the 130 patients who underwent dose adjustments during the LTE period, 116 (89.2%) had not discontinued participation in the study by week 96. By week 96, a total of 5 patients (3.8%) had discontinued participation in the study (3 patients [5.7%], 1 patient [2.5%], and 1 patient [2.7%] in the placebo group → ustekinumab q8w group, ustekinumab q12w group → ustekinumab q8w group, and ustekinumab q8w group → ustekinumab q8w group, respectively).

[0385] Demographic characteristics and baseline characteristics The baseline demographic and disease characteristics of the randomized population that underwent dose adjustments during the LTE period were largely consistent with the characteristics of the randomized population.

[0386] demographics The baseline demographic characteristics of randomized subjects who underwent dose adjustments during the LTE period were generally well-balanced across treatment groups. Overall, 62.3% were male, 75.4% were Caucasian, with a median age of 40.0 years and a median weight of 73.60 kg.

[0387] Clinical disease characteristics Disease characteristics at 44 weeks of maintenance The clinical disease characteristics at week 44 of randomized subjects who received LTE and underwent dose adjustment were generally similar between the ustekinumab q12w group → ustekinumab q8w group and the ustekinumab q8w group → ustekinumab q8w group (as shown below): ●Percentage of subjects in clinical remission (world definition): 40.0% and 51.4% ●Percentage of patients who underwent endoscopic treatment: 55.0% and 54.1% ● Average Mayo score: 2.9 and 2.8 ●Median IBDQ scores: 184.0 and 180.0 ●Median CRP concentration: 1.67 mg / L and 3.15 mg / L ●Median fecal calprotectin concentration: 152.0 mg / kg and 284.00 mg / kg ●Median fecal lactoferrin concentration: 15.36 μg / g and 24.22 μg / g

[0388] The clinical disease characteristics at week 44 in the placebo group, which underwent dose adjustment to ustekinumab q8w during the LTE period, were as follows: ●Percentage of subjects in clinical remission (world definition): 26.4% ●Percentage of patients who underwent endoscopic treatment: 37.7% ●Average Mayo score: 4.1 ●Median IBDQ score: 178.0 ●Median CRP concentration: 2.67 mg / L ●Median fecal calprotectin concentration: 726.50 mg / kg ●Median lactoferrin concentration in feces: 52.91 μg / g

[0389] Prior treatment and combination therapy Concomitant medications At baseline during the induction phase, 89.2% of patients in the maintenance phase who underwent dose adjustment with LTE received concomitant UC medication (88.7%, 85.0%, and 94.6% in the placebo → ustekinumab q8w group, ustekinumab q12w group → ustekinumab q8w group, and ustekinumab q8w group → ustekinumab q8w group, respectively). The overall proportions of patients receiving corticosteroids, immunomodulators, and aminosalicylates were 55.4%, 22.3%, and 70.0%, respectively. The proportions of patients receiving each type of UC medication in the placebo → ustekinumab q8w, ustekinumab q12w → ustekinumab q8w, and ustekinumab q8w → ustekinumab q8w groups were as follows: ● Corticosteroids: 49.1%, 60.0%, and 59.5% ● Immunomodulatory drugs: 26.4%, 17.5%, and 21.6% ● Aminosalicylates: 73.6%, 70.0%, and 64.9%

[0390] Medical history After dose adjustments were made via LTE, the majority of patients (95.4%) randomized to the maintenance phase either showed an inadequate response to or poor tolerance of corticosteroids and / or 6-MP / AZA at baseline during the induction phase, or exhibited corticosteroid dependence. Overall, 80.8% were refractory, dependent, or intolerant to corticosteroid administration, and 58.5% were refractory or intolerant to 6-MP / AZA administration.

[0391] Among the subjects randomized to the maintenance phase with dose adjustment via LTE, 41.5% had no history of biological agent failure at baseline during the induction phase (all were biological agent-naive). Among the 58.5% of subjects with a history of biological agent failure, the proportion of subjects in the overall dose adjustment group was similar (56.6%, 60.0%, and 59.5% in the placebo group → ustekinumab q8w group, ustekinumab q12w group → ustekinumab q8w group, and ustekinumab q8w group → ustekinumab q8w group, respectively).

[0392] Among those randomized during the maintenance phase, those who underwent dose adjustment with LTE were more likely to have a history of biological agent failure than the overall randomized population receiving LTE. The proportion of dose-adjusted patients with a history of biological agent failure was as follows: ● 57.7% were ineffective against at least one type of biological agent for anti-TNF (regardless of vedolizumab). -39.2% were unresponsive to biological agents targeting only anti-TNF (not vedolizumab). ● 19.2% of patients were unresponsive to vedolizumab (regardless of anti-TNF). ●18.5% were unresponsive to any anti-TNF and vedolizumab biological agents.

[0393] Deviation from the clinical trial protocol Between weeks 44 and 96, 27 patients (4.6%) experienced the following major deviations from the clinical trial protocol: ●Two cases (0.3%) were reported to have met the withdrawal criteria but did not withdraw. ● Fifteen cases (2.6%) were reported to have received incorrect administration or dosage. ● Eleven cases (1.9%) reported deviations from the clinical trial protocol for reasons other than those listed above (e.g., "other").

[0394] The subjects may be counted in two or more categories, or may have one or more deviations within a category.

[0395] Deviation from test drug administration Among those who received LTE, 15 cases of deviation from the study drug administration were reported. As described above, all 15 cases were classified as deviations from the primary clinical trial protocol.

[0396] Randomized subjects A total of 11 subjects (5 in the placebo group and 6 in the ustekinumab q12w group) were mistakenly re-administered ustekinumab q8w at their 56-week visit. Of the 5 subjects in the placebo group, 3 continued to receive the re-administered dose (ustekinumab q8w), while 2 were re-administered back to placebo. All subjects from the mistakenly re-administered ustekinumab q12w group were re-administered back to q12w at their subsequent visits.

[0397] In a total of two cases (both in the ustekinumab q12w group), the drug was not administered using the syringe assigned to the patient upon arrival, but instead using the wrong syringe (one patient was mistakenly given placebo upon arrival, and the other was mistakenly given ustekinumab instead of placebo upon arrival). The patients were reinstated to the dosage assigned to them at subsequent visits.

[0398] Non-randomized subjects Two patients were administered the wrong syringe instead of the one assigned to them at their visit. The patients were reinstated to their assigned dosage at subsequent visits. One patient from the placebo-induction responder group received an expired study drug at their 60-week visit. The patient was mistakenly given ustekinumab instead of placebo, and the ustekinumab administered was also expired. The patient was followed up for safety events, and no adverse events were reported. One patient from the delayed-induction responder group received a placebo instead of ustekinumab at their 56-week visit.

[0399] Meets the withdrawal criteria but does not withdraw. Of the subjects who received LTE, two (both from the non-randomized placebo-initiated responder group) were confirmed to have met the withdrawal criteria, but did not withdraw. Both subjects experienced a worsening of ulcerative colitis (UC) that did not improve after 16 weeks. However, one subject reported an adverse event (UC worsening AE) within the 16 weeks specified in the clinical trial protocol and discontinued administration of the study drug, while the other subject did not report an UC worsening AE during the LTE period, resulting in both deviations being reported incorrectly.

[0400] Deviation from unauthorized concomitant medications The administration of combination therapies was left to the discretion of the principal investigator, except for drugs explicitly prohibited in the clinical trial protocol. No subjects started taking prohibited drugs during the LTE period.

[0401] Other major deviations from clinical trial protocols Of the subjects who received LTE, 11 were classified as "other" and reported deviations from the clinical trial protocol.

[0402] Of the randomized subjects, 10 cases were reported to have "other" deviations from the clinical trial protocol: 2 in the placebo group and 4 each in the ustekinumab q12w and q8w groups. ●Among those in the placebo group: -One patient underwent local testing of serum ustekinumab levels and continued the trial. -In one case, clinical laboratory evaluations were not performed at weeks 56 and 68, and therefore the results could not be obtained by the next hospital visit for administration. Subsequent chemical and hematological test values ​​were within the normal range specified by the central laboratory. ●Among the subjects in the ustekinumab q12w group: -Four subjects received the test drug, which was later deemed unsuitable for use. All subjects were monitored for safety events after administration, but no adverse safety events were observed. ●Among the subjects in the ustekinumab q8w group: - In three cases, a urine pregnancy test was not performed at the time of the administration visit. Testing was resumed at subsequent visits, and none of the patients reported pregnancy. - In one case, a TB evaluation was not performed at the 60-week visit. A TB diagnosis was made at a subsequent visit, and no signs of active TB were observed.

[0403] Among the non-randomized participants, the only deviation from the clinical trial protocol classified as "other" was reported, as mentioned above, in one case from the placebo-initiated responder group who received an expired study drug at the 60-week visit.

[0404] Summary and impact of deviations from the clinical trial protocol Overall, among the 399 randomized and 189 non-randomized patients who received LTE, deviations from the study protocol were reported in 23 patients (5.8%) and 4 patients (2.1%), respectively. Most deviations (15 out of 27 patients) were classified as "incorrect administration or incorrect dose received." For patients who received an incorrect dose of ustekinumab, the analysis was performed on the assigned treatment group, including those with dose adjustment errors. For patients in the placebo group who received confirmed ustekinumab, it was assumed that dose adjustment had been performed, and safety was analyzed in the ustekinumab q8w group from the time the patient received ustekinumab. Deviations related to procedures specified in the study protocol (i.e., investigational drug monitoring, urine pregnancy testing, TB risk assessment) were addressed by the institutions, and their impact on patient safety was examined. No safety issues were identified.

[0405] During the trial period, specific deviations were addressed at the facility level and through facility-wide communication and training. Regarding issues during the trial, appropriate corrective and preventive measures were taken before the DBL (Discipline-Based Learning) phase.

[0406] In summary, deviations from the clinical trial protocol were of varying natures and were determined not to have a clinically relevant impact on data integrity or patient safety. Similarly, no significant impact of deviations was observed on the safety profile of ustekinumab in this study. Overall, the observed safety profile was consistent with the labeled safety information for ustekinumab in the disease under study and for other indications.

[0407] Treatment compliance Administration of the study drug was performed by appropriately licensed healthcare professionals according to the dosing groups assigned by the IWRS. Compliance with dosing assignments was managed by site personnel. Site personnel administered the study drug and recorded the amount administered. All subjects' eCRFs were monitored by site monitors designated by the sponsor. During these monitoring visits, all procedures were evaluated for compliance with the study protocol. Missed study visits were recorded in the eCRF. Site monitors designated by the sponsor verified source documents, ensured accountability for the study drug, and ensured site-wide compliance. Subject records were reviewed and compared with eCRF data entries to verify consistency. The study drug was not to be used for purposes other than those outlined in the study protocol. Used study drug vials and syringes were stored at the site until site monitors checked the study drug accountability forms.

[0408] Degree of exposure In total, 454 patients received at least one dose of ustekinumab during the LTE period. ● Placebo: Placebo was administered to 188 patients (ustekinumab dose: 0.0 mg). ●90mg q12w: 141 patients received a median cumulative dose of 450.0mg. ●90mg q8w: A median cumulative dose of 630.0mg was administered to 353 patients.

[0409] In the maintenance phase, participants who were randomized and received LTE were administered the following study drugs from week 44 to week 96, or until dose adjustment: ● Placebo: Placebo was administered to 115 patients (ustekinumab dose: 0.0 mg). ●90mg q12w: 141 patients received a median cumulative dose of 450.0mg. ●90mg q8w: 143 patients received a median cumulative dose of 630.0mg.

[0410] Among the ustekinumab delayed response group, a total of 116 patients who received LTE (ustekinumab 90 mg SC q8w administration) received a median cumulative dose of 630.0 mg from week 44 to week 96.

[0411] Of the placebo-initiated responders, a total of 73 patients continued to receive placebo via LTE. One patient in this group received ustekinumab. Safety data prior to ustekinumab administration for this patient were included in the placebo group, and safety data after ustekinumab administration were included in the ustekinumab q8w group as appropriate.

[0412] The degree of exposure in subjects who underwent dose adjustment. Among those randomized to ustekinumab during the maintenance phase, those whose dose was adjusted during the LTE period received ustekinumab from the dose adjustment until week 96 as follows: ● Placebo → Ustekinumab q8w: 53 patients received a median cumulative dose of 450.0 mg. ● Ustekinumab q12w → Ustekinumab q8w: A median cumulative dose of 270.0 mg was administered to 40 patients. ● Ustekinumab q8w → Ustekinumab q8w: 37 patients were administered a cumulative median dose of 180.0 mg.

[0413] Pharmacokinetic and immunogenicity results Pharmacokinetics All patients who received at least one dose of ustekinumab during the LTE period were included in the PK analysis. For randomized subjects, serum ustekinumab concentrations up to the time of dose adjustment were aggregated. For randomized subjects who underwent dose adjustment, an overview of ustekinumab concentrations from the time of dose adjustment onward is provided. For non-randomized subjects, ustekinumab concentration data were aggregated for delayed responders of ustekinumab induction. Results from week 44 to week 92 are aggregated in this report, and results from week 0 to week 44 are shown in UCO3001 44W.

[0414] Of the subjects randomized during the maintenance phase, a total of 337 continued to receive ustekinumab during the LTE (Long Life Stage). This included 141 patients receiving ustekinumab 90 mg SC q12w, 143 patients receiving ustekinumab 90 mg SC q8w, and 53 placebo patients whose dose was adjusted to ustekinumab q8w during the LTE period. Of the 141 randomized patients who received ustekinumab q12w during the LTE, 40 underwent dose adjustment to ustekinumab q8w. Of the 143 randomized patients who received ustekinumab q8w during the LTE, 37 underwent sham dose adjustment (i.e., continued ustekinumab q8w).

[0415] Randomized subjects Based on the trial visit schedule, blood samples for measuring serum ustekinumab concentrations were collected every 24 weeks starting from week 44 (i.e., at weeks 44, 68, and 92). Therefore, concentration data for subjects who received ustekinumab at q12w until dose adjustment were obtained 8 weeks after ustekinumab administration at weeks 36, 60, and 84, but not in the trough. On the other hand, concentration data for subjects who received ustekinumab at q8w were obtained 4 weeks after ustekinumab administration at weeks 40, 64, and 88, but not in the trough.

[0416] Randomized subjects who received ustekinumab during the LTE period maintained their ustekinumab concentration throughout the LTE period. At 44 weeks after the start of LTE (equivalent to 8 weeks after the last maintenance dose in the ustekinumab q12w group and 4 weeks after the last maintenance dose in the ustekinumab q8w group), the median [mean] serum ustekinumab level was approximately three times higher in the ustekinumab q8w group (9.41 [8.84] μg / mL) than in the q12w group (2.50 [3.02] μg / mL). Among subjects randomized to ustekinumab q12w during the maintenance phase, those who continued administration of ustekinumab 90 mg via LTE (i.e., weeks 48, 60, 72, and 84) had a median ustekinumab concentration of 2.13 μg / mL to 2.59 μg / mL during the 8-week period after ustekinumab administration (weeks 68 to 92). Among subjects randomized to ustekinumab q8w during the maintenance phase, those who continued administration of ustekinumab 90 mg via LTE (i.e., weeks 48, 56, 64, 72, 80, and 88) had a median ustekinumab concentration of 6.38 μg / mL to 6.65 μg / mL during the 4-week period after ustekinumab administration (weeks 68 to 92).

[0417] [Table 9]

[0418] These results indicate that randomized subjects who continued administration of either the ustekinumab q12w or q8w regimen in the LTE maintained sustained and consistent ustekinumab concentrations up to week 92 of the LTE, which were generally comparable to the serum ustekinumab concentrations observed during the study maintenance phase.

[0419] Randomized subjects who underwent dose adjustment In maintenance trials, patients whose UC disease activity worsened during the LTE period were randomized and received one dose adjustment starting at week 56. Three scenarios were considered for those who met the criteria for dose adjustment: ●Participants randomized to placebo received a dose adjustment to ustekinumab 90 mg q8w. ● Patients randomized to ustekinumab 90 mg q12w received a dose adjustment to ustekinumab 90 mg q8w. ● Patients randomized to ustekinumab 90 mg q8w continued with the same dosing regimen (sham dose adjustment).

[0420] Because the subjects who underwent dose adjustments initiated ustekinumab q8w at different hospital visits, the summary of concentration data for these subjects does not represent expected concentrations over time relative to ustekinumab q8w. However, as expected, serum ustekinumab concentrations increased after dose adjustment from placebo to ustekinumab q8w. Specifically, the median serum ustekinumab concentration increased from 0.0 μg / mL at week 44 to 4.70 μg / mL at week 68 and to 3.64 μg / mL at week 92.

[0421] In patients randomized to ustekinumab q12w, serum ustekinumab concentrations at week 44 were similar between those who received dose adjustment and those who did not (2.51 μg / mL and 2.50 μg / mL, respectively). After dose adjustment from ustekinumab q12w to ustekinumab q8w, the median serum concentrations at weeks 68 and 92 were 2.97 μg / mL and 3.83 μg / mL, respectively.

[0422] Among the subjects randomized to the ustekinumab q8w group, the median ustekinumab concentration from week 44 to week 92 was approximately the same between subjects who underwent dose adjustment and those who did not.

[0423] Non-randomized group (ustekinumab delayed response patients) Delayed response patients to ustekinumab induction were those who did not respond to ustekinumab IV induction at week 0, received ustekinumab 90 mg SC at week 8 of the induction period, and showed a clinical response at week 16 of the induction period. These patients continued to receive ustekinumab 90 mg SC at q8w in the maintenance study and LTE.

[0424] At week 44, the median serum concentration (7.83 μg / mL) in the delayed response group of ustekinumab induction was slightly lower than that of the group randomized to ustekinumab q8w in response to a single ustekinumab IV induction dose and without dose adjustment (9.67 μg / mL). This difference in ustekinumab concentrations disappeared at weeks 68 and 92, and the median concentrations of delayed responders (6.21 μg / mL and 5.94 μg / mL, respectively) were comparable to those of the group randomized to ustekinumab q8w and without dose adjustment (6.49 μg / mL and 6.66 μg / mL, respectively).

[0425] immunogenicity Immunogenicity (antibodies against ustekinumab) analysis was performed on all patients who received ustekinumab and on randomized subjects. The relationship between antibodies against ustekinumab and serum ustekinumab concentrations in the randomized subjects was also examined.

[0426] After ustekinumab administration, the expression rate of antibodies against ustekinumab remained low until week 96 of LTE.

[0427] Immunogenicity up to 96 weeks Subjects who received ustekinumab initiation, maintenance, and long-term continuation therapy. In a group of 400 patients who received ustekinumab during the maintenance phase and continued ustekinumab administration during the late-stage therapy (LTE) (this group consisted of subjects who achieved a clinical response after induction of ustekinumab IV and were randomized to ustekinumab SC during the maintenance phase, and subjects whose response was delayed at week 16 of the induction phase and who subsequently received SC maintenance therapy), 22 patients (5.5%) were positive for antibodies against ustekinumab between week 0 and week 96 of the LTE induction phase. The expression rate of antibodies against ustekinumab in this control group is considered to be the most relevant because it reflects the way ustekinumab is used in clinical practice. Most of the subjects who were positive for antibodies against ustekinumab (18 out of 22) had titers of ≤1:800, and 4 out of 22 patients (18.2%) were positive for neutralizing antibodies (NAb).

[0428] Randomized subjects A total of 399 randomized patients in the maintenance phase (284 receiving ustekinumab, 115 receiving placebo) received administration during the LTE period, and appropriate samples were collected at some point up to week 96 to assess antibody status against ustekinumab. The overall antibody expression rate against ustekinumab among the randomized patients was 6.8% (27 out of 399 patients). Among patients who did not undergo dose adjustment (including sham adjustment) during the LTE, the antibody expression rate against ustekinumab was similar between those who received ustekinumab for 12 weeks (5.0%) and those who rec...

Claims

1. A pharmaceutical composition for use in a method for treating moderate to severe active ulcerative colitis (UC) in a subject in need thereof, comprising: The pharmaceutical composition comprises an anti-IL-12 / IL-23p40 antibody, the antibody comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising a complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprising a complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRL3 amino acid sequence of SEQ ID NO: 6; The method includes administering to the subject the pharmaceutical composition; the antibody is administered to the subject intravenously at a dose of 130 mg, 260 mg, 390 mg, or 520 mg per administration at week 0 of treatment, and subcutaneously to the subject at a dose of 90 mg per administration at week 8 of treatment, and the antibody is administered at a maintenance dose every 8 weeks after week 8 of treatment or every 12 weeks after week 8 of treatment; The pharmaceutical composition is for use in treating a subject comprising: (a) symptomatic remission, (b) partial Mayo remission; (c) Mayo rectal bleeding subscore of 0; (d) Mayo stool frequency subscore of 0 or 1; (e) a decrease in mean absolute fecal count of at least 3; (f) reduction in corticosteroid use and / or dosage; (g) corticosteroid-free symptomatic remission; (h) Corticosteroid-free partial Mayo remission; (i) normalization of fecal lactoferrin; (j) normalization of fecal calprotectin levels; (k) Inflammatory Bowel Disease Questionnaire (IBDQ) total score improved by 16 points or more from the baseline of the induction phase; (l) IBDQ remission; (m) SF-36 PCS score improvement of 5 points or more from baseline in the induction phase; and (n) SF-36 MCS score improved by 5 points or more from baseline in the induction phase and (iii) being a responder to treatment at 92 weeks based on meeting one or more clinical endpoints selected from the group consisting of: The pharmaceutical composition.

2. The pharmaceutical composition of claim 1 , wherein the antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO:

8.

3. The pharmaceutical composition described in claim 1, wherein the antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO: 10 and a light chain having the amino acid sequence of SEQ ID NO:

11.

4. 4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition for intravenous administration further comprises a solution comprising 10 mM L-histidine, 8.5% (w / v) sucrose, 0.04% (w / v) polysorbate 80, 0.4 mg / mL L-methionine, and 20 μg / mL EDTA disodium salt dihydrate at pH 6.

0.

5. 5. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition for subcutaneous administration further comprises a solution comprising 6.7 mM L-histidine, 7.6% (w / v) sucrose, 0.004% (w / v) polysorbate 80 at pH 6.

0.

6. The pharmaceutical composition of claim 4, wherein the subject is a delayed induction responder.

7. 5. The pharmaceutical composition of claim 4, wherein the subject has previously failed or been intolerant to at least one therapy selected from the group consisting of anti-TNF, vedolizumab, corticosteroids, azathioprine (AZA), and 6-mercaptopurine (6MP), or the subject has demonstrated corticosteroid dependence.

8. 5. The pharmaceutical composition of claim 4, wherein the subject is identified as having mucosal healing that continues at least 92 weeks after week 0.

9. 1. A pharmaceutical composition comprising an anti-IL-12 / IL-23p40 antibody for treating moderate to severely active ulcerative colitis (UC) in a subject in need thereof, wherein the antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprises a complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRL3 amino acid sequence of SEQ ID NO: 6; The pharmaceutical composition is for use in treating a subject comprising: (a) symptomatic remission, (b) partial Mayo remission; (c) Mayo rectal bleeding subscore of 0; (d) Mayo stool frequency subscore of 0 or 1; (e) a decrease in mean absolute fecal count of at least 3; (f) reduction in corticosteroid use and / or dosage; (g) corticosteroid-free symptomatic remission; (h) Corticosteroid-free partial Mayo remission; (i) normalization of fecal lactoferrin; (j) normalization of fecal calprotectin levels; (k) Inflammatory Bowel Disease Questionnaire (IBDQ) total score improved by 16 points or more from the baseline of the induction phase; (l) IBDQ remission; (m) SF-36 PCS score improvement of 5 points or more from baseline in the induction phase; and (n) SF-36 MCS score improved by 5 points or more from baseline in the induction phase and (iii) being a responder to treatment at 92 weeks based on meeting one or more clinical endpoints selected from the group consisting of: The pharmaceutical composition.

10. The pharmaceutical composition of claim 9 , wherein the antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO:

8.

11. The pharmaceutical composition of claim 9 , wherein the antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO: 10 and a light chain having the amino acid sequence of SEQ ID NO: 11.

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