IL-23 SPECIFIC ANTIBODIES FOR THE TREATMENT OF SYSTEMIC SCLEROSIS - Patent application

Abstract

A method of treating systemic sclerosis in a patient comprises administering an IL-23 specific antibody, e.g., guselkumab, in an initial subcutaneous dose and subsequent subcutaneous doses, such that the patient responds to the antibody and meets one or more clinical endpoints.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) (Reference to electronically submitted sequence listing) This application contains a Sequence Listing that has been submitted electronically through the U.S. Patent and Trademark Office Patent Center as an XML format Sequence Listing with the file name "JBI6681WOPCT1 Sequence Listing.xml", created on December 12, 2022, and 11 kb in size. The Sequence Listing submitted through the Patent Center is a part of this specification and is incorporated by reference in its entirety herein.

[0002] FIELD OF THEINVENTION The present invention is directed to methods of treating systemic sclerosis using antibodies that bind to human IL23. In particular, the present invention relates to doses and dosing regimens and uses and methods for administering anti-IL23 specific antibodies and certain pharmaceutical compositions of the antibodies. [Background technology]

[0003] Systemic sclerosis (SSc) is a complex autoimmune disease characterized by thickening and hardening of the skin and visceral (gastrointestinal, heart, lungs, and kidney) involvement with a chronic and often progressive course. One of the clinical features of SSc is its interpatient variability, with heterogeneity observed in clinical manifestations, autoantibody profiles, disease progression, response to treatment, and survival. Based on the extent of their skin involvement, patients are classified into limited cutaneous SSc (lcSSc) and diffuse cutaneous SSc (dcSSc) subsets. In lcSSc, dermal fibrosis is limited to the fingers (sclerodactyly), distal extremities, and face. On the other hand, patients with dcSSc have dermal fibrosis in areas including the trunk and proximal extremities, relatively rapid disease progression with widespread skin changes, and early onset of visceral involvement compared to lcSSc. Systemic sclerosis is designated as an intractable disease in Japan, and according to the Japan Rare Diseases Information Center, there were 26,740 patients with moderate to severe SSc in 2018. Worldwide, the prevalence of SSc ranges from 7 to 489 cases per million, and the incidence ranges from 0.6 to 122 cases per million annually.

[0004] Systemic sclerosis has a high disease burden and unmet medical need, as most of the current treatments have limited efficacy. No drugs significantly affect the natural history of SSc, and no advanced therapies have been approved for the treatment of SSc in its entirety. Clinical trials of autologous hematopoietic stem cell transplantation (HSCT) in dcSSc have demonstrated survival benefits, including meaningful improvements in skin and lung fibrosis, and health-related quality of life. However, these benefits must be weighed against the increased risk of transplant-related mortality, which limits the wider use of HSCT in the treatment of SSc.

[0005] Survival rates have improved in recent decades and correlate best with clinical disease subtype (dcSSc vs. lcSSc) and the degree of organ involvement. The 5-year survival rate of patients with dcSSc improved significantly from 69% in the 1990-1993 cohort to 84% in the 2000-2003 cohort. The 5-year survival rate of patients with lcSSc has remained at a very high level and did not change over the same time periods (93% and 91%, respectively).

[0006] Mortality associated with scleroderma renal crisis has declined significantly over the last few decades with the use of angiotensin-converting enzyme inhibitors. In contrast, pulmonary complications (ILD and / or pulmonary arterial hypertension) remain the most common cause of death in patients with SSc. In contrast to rheumatoid arthritis (RA), the concept and use of disease-modifying therapies to attenuate or reverse pathology and clinical effects has not currently been applied to SSc.

[0007] Various drugs are used to treat specific symptoms or organ systems. Low-dose corticosteroids may be useful for skin sclerosis in patients with acute progression, but are used only when necessary as they may predispose to renal crisis. Nintedanib, a tyrosine kinase inhibitor, is approved for the treatment of ILD associated with SSc. Various immunosuppressants, including methotrexate, azathioprine, mycophenolate mofetil, and cyclophosphamide, may help with alveolitis. Mycophenolate mofetil is also effective in treating ILD. Prostacyclin and endothelin receptor antagonists are used for pulmonary hypertension. Calcium channel blockers may help with Raynaud's phenomenon. Intravenous (IV) infusion of prostaglandin E1 or prostacyclin or sympatholytic drugs can be used for digital ischemia. Reflux esophagitis is relieved by proton pump inhibitors.

[0008] Although various agents are used to treat some of the symptoms of SSc, there remains a high unmet medical need for disease-modifying therapies with defined efficacy and safety profiles for SSc.

[0009] Interleukin (IL)-12 is a secreted heterodimeric cytokine composed of two disulfide-linked glycosylated protein subunits (designated p35 and p40 for their approximate molecular weight). IL-12 is primarily produced by antigen-presenting cells and promotes cell-mediated immunity by binding to a two-chain receptor complex expressed on the surface of T cells or natural killer (NK) cells. The IL-12 receptor beta-1 (IL-12Rβ1) chain binds to the p40 subunit of IL-12, providing the primary interaction between IL-12 and its receptor. However, it is IL-12p35 ligation of the second receptor chain, IL-12Rβ2, that confers intracellular signaling (e.g., STAT4 phosphorylation) and activation of receptor-bearing cells (Presky et al, 1996). IL-12 signaling, concomitant with antigen presentation, is believed to trigger T cell differentiation towards the T helper 1 (Th1) phenotype, characterized by interferon gamma (IFNγ) production (Trinchieri, 2003). Th1 cells are believed to promote immunity against several intracellular pathogens, generate complementary antibody isotypes, and contribute to tumor immunosurveillance. Therefore, IL-12 is believed to be a critical component of the host defense immune mechanism.

[0010] It was discovered that the p40 protein subunit of IL-12 can also associate with a separate protein subunit designated p19 to form a novel cytokine, IL-23 (Oppman et al, 2000). IL-23 also signals through a two-chain receptor complex. As the p40 subunit is shared between IL-12 and IL-23, it follows that the IL-12Rβ1 chain is also shared between IL-12 and IL-23. However, it is IL-23p19 ligation of the second component of the IL-23 receptor complex, IL-23R, that confers IL-23-specific intracellular signaling (e.g., STAT3 phosphorylation) and subsequent IL-17 production by T cells (Parham et al, 2002; Aggarwal et al, 2003). Recent studies have shown that the biological functions of IL-23 and IL-12 are distinct, despite the structural similarities between these two cytokines (Langrish et al, 2005).

[0011] Abnormal regulation of IL-12 and Th1 cell populations has been implicated in many immune-mediated diseases, as neutralization of IL-12 with antibodies has been effective in treating animal models of psoriasis, multiple sclerosis (MS), rheumatoid arthritis, inflammatory bowel disease, insulin-dependent (type 1) diabetes mellitus, and uveitis (Leonard et al., 1995; Hong et al., 1999; Malfait et al., 1998; Davidson et al., 1998). However, these studies neutralized both IL-12 and IL-23 in vivo, as they targeted the common p40 subunit. Thus, it is unclear whether IL-12 or IL-23 mediated the disease, or whether both cytokines need to be inhibited to achieve disease suppression. Recent studies have confirmed that IL-23 inhibition may offer benefits comparable to anti-IL-12p40 strategies using IL-23p19-deficient mice or specific antibody neutralization of IL-23 (Cua et al., 2003; Murphy et al., 2003; Benson et al., 2004).

[0012] Although the pathogenesis of SSc is complex and multifactorial, accumulating evidence suggests that the IL-23 / IL-17 pathway is involved in the pathophysiology of SSc. Several clinical studies have shown that serum IL-23 / IL-17 and skin IL-23 are increased in SSc patients compared with healthy volunteers. Serum IL-23 levels and A correlation with the severity / extent of interstitial pulmonary involvement was observed in patients with SSc.

[0013] A case report showed that ustekinumab, a mAb against IL-12 and IL-23, successfully improved skin thickening in a patient with both psoriasis and SSc. In Japan, a phase 3 clinical trial is underway to evaluate brodalumab, an IL-17R antibody, in subjects with SSc. Results of a prior brodalumab phase 1b trial (n=8) (NCT04368403) are not publicly available, but an ongoing phase 3 trial has been initiated to evaluate efficacy and safety outcomes from the phase 1 trial.

[0014] Taken together, these observations provide sufficient rationale to investigate the function of IL-23 in the pathogenesis of SSc in this proof-of-concept (PoC) trial of guselkumab in participants with SSc.

[0015] In summary, there remains a substantial unmet medical need for new treatment options for SSc, particularly therapies with novel mechanisms of action that have the potential to raise the efficacy bar and maximize the proportion of patients who achieve and maintain clinical remission. Summary of the Invention [Means for solving the problem]

[0016] In a first aspect, the present invention relates to a method of treating a subject (patient) suffering from SSc comprising administering an anti-IL23 specific antibody (also called IL23p19 or IL23p19 subunit antibody), such as guselkumab, with a first dose at the start of treatment and at time intervals thereafter, preferably about every 4 weeks after the first dose, such as administration at weeks 0, 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and / or 48. Moreover, in another embodiment, the treatment continues for 96 weeks or more after initiation of treatment.

[0017] In another aspect, the invention relates to the use of an anti-IL23 specific antibody, such as guselkumab, for the treatment of SSc in a subject, wherein the antibody is administered as an initial dose at the start of treatment and at subsequent time intervals, preferably about every 4 weeks after the initial dose, such as at weeks 0, 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and / or 48. Moreover, in another embodiment, the treatment continues for 96 weeks or more after initiation of treatment.

[0018] In one embodiment, the subject receives an anti-IL23 specific antibody intravenously at (i) an initial dose of 400 mg, 4 weeks after the initial dose, and 8 weeks after the initial dose, and (ii) continues treatment with subcutaneous administration of the anti-IL23 specific antibody at doses of 200 mg every 4 weeks thereafter.

[0019] In another embodiment, the composition used in the methods of the invention comprises a pharmaceutical composition comprising an anti-IL23 specific antibody.

[0020] In one embodiment, the SSc patient achieves a significant improvement in a clinical and / or exploratory endpoint selected from the group consisting of: (i) change from baseline in mRSS; (ii) worsening of mRSS; (iii) achieving an American College of Rheumatology Combined Response Index (ACR CRISS) score of 0.6 in dcSSc; (iv) change from baseline in forced vital capacity (FVC) and percent of predicted FVC; (v) Change from baseline in the measured absolute diffusing capacity of the lung for carbon monoxide (DLCO) and the derived percent predicted DLCO; (vi) change from baseline in number of digital ulcers in patients with digital ulcers at baseline; (vii) change from baseline in Health Assessment Questionnaire-Disability Index (HAQ-DI) score; (viii) change from baseline in the Frequency Scale for the Symptoms of Gastroesophageal Reflux Disease (FSSG) score; (ix) change from baseline in fibrotic changes and worsening of fibrotic changes as assessed by high-resolution computed tomography (HRCT); and (x) Change from baseline in capillaroscopic assessment in Patient global assessment (PGA) and / or Physician global assessment (PhGA).

[0021] In preferred embodiments, the endpoint may be measured at about 24, 52, or 104 weeks after initial treatment.

[0022] In another aspect of the invention, a pharmaceutical composition comprises an isolated anti-IL23 specific antibody having CDR sequences comprising: (i) heavy chain CDR amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and (ii) light chain CDR amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, optionally present in a composition of 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80 of the pharmaceutical composition, and the diluent is standard water.

[0023] Another aspect of the method of the invention comprises administering said pharmaceutical composition comprising an isolated anti-IL23 specific antibody having a heavy chain variable region amino acid sequence of SEQ ID NO:7 and a light chain variable region amino acid sequence of SEQ ID NO:8, optionally present in a pharmaceutical composition of 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80, and the diluent is standard water for use in treating patients with SSc according to the doses and dosing regimens described herein.

[0024] A further aspect of the method of the invention comprises administering said pharmaceutical composition comprising an isolated anti-IL23 specific antibody having a heavy chain amino acid sequence of SEQ ID NO:9 and a light chain amino acid sequence of SEQ ID NO:10, optionally present in the following composition of pharmaceutical composition: 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80, wherein the diluent is standard water.

[0025] In yet a further embodiment, the method of the invention comprises administering a pharmaceutical composition comprising the antibody guselkumab (sold as Tremfya® by Janssen Biotech, Inc.), optionally in the following composition of the pharmaceutical composition: 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80, and the diluent is standard water.

[0026] The details of one or more embodiments of the invention are set forth in the description below. Other features and advantages will become apparent from the following detailed description, the drawings, and the appended claims. [Brief description of the drawings]

[0027] [Figure 1] FIG. 1 shows a schematic diagram of the main trial described herein. Abbreviations: DBL = database locked; IV = intravenous; N = number of participants; PE = primary endpoint; R = randomized; SC = subcutaneous; SE = secondary endpoint. [Diagram 2] 1 shows a schematic diagram of the LTE test described herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] As used herein, methods of treating a subject suffering from SSc include administering isolated, recombinant, and / or synthetic anti-IL-23 specific human antibodies, as well as diagnostic and therapeutic compositions, methods, and devices.

[0029] As used herein, "anti-IL-23 specific antibodies," "anti-IL-23 antibodies," "antibody portions," or "antibody fragments," and / or "antibody variants," and the like, include any protein or peptide-containing molecule comprising at least a portion of an immunoglobulin molecule, such as, but not limited to, at least one complementarity determining region (CDR) or ligand binding portion thereof of a heavy or light chain, a heavy or light chain variable region, a heavy or light chain constant region, a framework region, or any portion thereof, or at least a portion of an IL-23 receptor or binding protein, that can be incorporated into an antibody of the invention. Such antibodies optionally further affect a specific ligand, for example, but not limited to, such antibodies modulate, reduce, enhance, antagonize, stimulate, reduce, ameliorate, block, inhibit, abrogate, and / or prevent at least one IL-23 activity or binding, or IL-23 receptor activity or binding, in vitro, in situ, and / or in vivo. As non-limiting examples, suitable anti-IL-23 antibodies, specified portions, or variants of the invention can bind to at least one IL-23 molecule or a specified portion, variant, or domain thereof. Suitable anti-IL-23 antibodies, specified portions, or variants can also, optionally, affect at least one IL-23 activity or function, including, but not limited to, RNA, DNA, or protein synthesis, IL-23 release, IL-23 receptor signaling, membrane IL-23 cleavage, IL-23 activity, IL-23 production, and / or synthesis.

[0030] The term "antibody" is further intended to encompass antibodies, digested fragments thereof, specific portions, and variants thereof, including antibody mimetics or portions of antibodies that mimic the structure and / or function of antibodies, such as single chain antibodies and fragments thereof, or specific fragments or portions thereof. Functional fragments include antigen-binding fragments that bind to mammalian IL-23. For example, antibody fragments capable of binding to IL-23 or portions thereof, including but 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 biology techniques) fragments are encompassed by the present invention (see, e.g., Colligan, Immunology, supra).

[0031] Such fragments can be produced by enzymatic cleavage, synthetic, or recombinant techniques known in the art and / or described herein. Antibodies can also be produced in a variety of truncated forms using antibody genes in which one or more stop codons have been introduced upstream of the natural stop site. For example, a combination of genes encoding a F(ab')2 heavy chain portion can be used to encode the C(ab')2 heavy chain portion. H The antibody can be designed to contain DNA sequences encoding the antibody domain and / or hinge region. The various portions of the antibody can be joined chemically by conventional techniques, or can be prepared as a contiguous protein using genetic engineering techniques.

[0032] As used herein, the term "human antibody" refers to an antibody that is human and contains substantially all parts of the protein (e.g., CDRs, frameworks, C L , C H Domain (e.g., C H 1. C H 2. C H 3), Hinge (V L , V H)) that are substantially non-immunogenic in humans with only minor sequence changes or mutations. A "human antibody" may be an antibody derived from or closely corresponding to a human germline immunoglobulin sequence. A human antibody may contain amino acid residues not encoded by a germline immunoglobulin sequence (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). In many cases, this means that the human antibody is substantially non-immunogenic in humans. Human antibodies have been classified into groups based on the similarity of their amino acid sequences. Thus, using sequence similarity searches, antibodies with similar linear sequences can be selected as templates for creating human antibodies. Similarly, antibodies with names including primates (monkeys, baboons, chimpanzees, etc.), rodents (mouse, rats, rabbits, guinea pigs, hamsters, etc.), and other mammals designate antibodies specific for such species, subgenus, genus, subfamily, and family. Furthermore, chimeric antibodies can include any combination of the above. Such changes or mutations optionally and preferably retain or reduce immunogenicity in humans or other species compared to the unmodified antibody. Thus, a human antibody is distinct from a chimeric antibody or a humanized antibody.

[0033] It is noted that human antibodies can be produced by non-human animals or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. Furthermore, when the human antibody is a single-chain antibody, it can contain a linker peptide not found in naturally occurring human antibodies. For example, an Fv can contain a linker peptide, such as 2 to about 8 glycine or other amino acid residues, connecting the variable region of the heavy chain and the variable region of the light chain. Such a linker peptide is considered to be of human origin.

[0034] Bispecific, heterospecific, heterobinding or similar antibodies may also be used, which are monoclonal, preferably human or humanized, antibodies with binding specificities for at least two different antigens, one of which is for at least one IL-23 protein and the other for any other antigen. Methods for producing bispecific antibodies are known in the art. Traditionally, recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-light chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature 305:537 (1983)). Due to the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a possible mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule (usually done by affinity chromatography steps) is quite laborious and the product yield is low. Similar procedures are described, for example, in WO 93 / 08829, U.S. Pat. Nos. 6,210,668, 6,193,967, 6,132,992, 6,106,833, 6,060,285, 6,037,453, 6,010,902, 5,989,530, 5,959,084, 5,959,083, 5,989,530, 5,959,084, 5,959,083, 5,989,530, 5,959,084, 5,959,085, 5,989,530 ...89,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,530, 5,989,5 Nos. 32448, 5833985, 5821333, 5807706, 5643759, 5601819, 5582996, 5496549, 4676980, WO 91 / 00360, WO 92 / 00373, EP 03089, Traunecker et al., EMBO J. 10:3655 (1991), and Suresh et al., Methods in Enzymology 121:210 (1986), each of which is incorporated herein by reference in its entirety.

[0035] Anti-IL-23 specific antibodies (also referred to as IL-23 specific antibodies) (or antibodies against IL-23) useful in the methods and compositions of the invention may optionally be characterized by high affinity binding to IL-23, and optionally and preferably low toxicity. In particular, antibodies of the invention, specific fragments or variants thereof, in which the individual components such as the variable region, constant region and framework, individually and / or collectively, optionally and preferably have low immunogenicity, are useful in the present invention. Antibodies that can be used in the present invention are optionally characterized by the ability to treat patients for extended periods of time with measurable alleviation of symptoms and low and / or acceptable toxicity. Low or acceptable immunogenicity and / or high affinity, as well as other favorable properties, can contribute to the therapeutic results obtained. "Low immunogenicity" is defined herein as raising a significant HAHA, HACA, or HAMA response in less than about 75%, or preferably less than about 50%, of treated patients, and / or raising low titers (less than about 300, preferably less than about 100, as measured by double antigen enzyme immunoassay) in treated patients (Elliott et al., Lancet 344:1125-1127 (1994), incorporated herein by reference in its entirety). "Low immunogenicity" can also be defined as the incidence of titrable levels of antibodies to an anti-IL-23 antibody in patients treated with an anti-IL-23 antibody when occurring in less than 25% of treated patients, preferably less than 10% of treated patients, at the recommended dose over the recommended course of treatment during the treatment period.

[0036] The term "safety", when referring to an administration, dosing regimen, treatment or method with an anti-IL-23 antibody of the present invention (e.g., the anti-IL-23 antibody guselkumab), refers to a relatively low or reduced frequency and / or a low or reduced severity of treatment-emergent adverse events (also referred to as AEs or TEAEs), e.g., from an conducted clinical trial, e.g., a Phase 2 clinical trial, and earlier clinical trials, compared to a standard of care or another comparison standard. An adverse event is an untoward medical occurrence in a patient administered a medicinal product. In particular, when referring to an administration, dosing regimen or treatment with an anti-IL-23 antibody of the present invention, clinically proven safety refers to a relatively low or reduced frequency and / or a low or reduced severity of adverse events associated with administration of the antibody, when the cause is considered possible, probable, or highly likely to be due to the use of the anti-IL-23 antibody.

[0037] usefulness The isolated nucleic acids of the invention can be used to produce at least one anti-IL-23 antibody or specific variants thereof, which can be used to measure or affect cells, tissues, organs, or animals (including mammals and humans) to diagnose, monitor, regulate, treat, alleviate, help prevent the occurrence of, or reduce the symptoms of SSc.

[0038] Such methods may include administering to a cell, tissue, organ, animal, or patient in need of such modulation, treatment, amelioration, prevention, or reduction of a symptom, effect, or mechanism an effective amount of a composition or pharmaceutical composition comprising at least one anti-IL-23 antibody. An effective amount may include an amount of about 0.001-500 mg / kg per single (e.g., bolus), multiple, or continuous administration, or an amount that achieves a serum concentration of 0.01-5000 μg / mL per single, multiple, or continuous administration, or any effective range or value therein, performed and determined using known methods described herein or known in the relevant art.

[0039] Cited literature All publications or patents cited herein, whether specifically stated or not, are incorporated herein by reference in their entirety, indicating the prior art at the time of the invention and / or providing a description and enabling of the invention. A publication refers to any scientific publication or patent publication or any other information available in any media format, including all recorded in electronic or printed form. The following documents are incorporated herein by reference in their entirety: 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, 2 nd 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), Colligan et al., Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001).

[0040] Antibodies of the invention - production and purification At least one anti-IL-23 used in the methods of the invention can optionally be produced by a cell line, mixed cell line, immortalized cell, or clonal population of immortalized cells, as known in the art, see, for example, 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, 2002; and, 2003, Immunotherapy and Immunotherapy, 1999, 144:1311-1320, each of which is incorporated herein by reference in its entirety.nd 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), Colligan et al., Current Protocols in Protein Science, John Wiley & See Sons, NY, NY, (1997-2001).

[0041] A preferred anti-IL-23 antibody is guselkumab (also known as CNTO1959), which has a heavy chain variable region amino acid sequence of SEQ ID NO:7 and a light chain variable region amino acid sequence of SEQ ID NO:8, and heavy chain CDR amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and light chain CDR amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6. Other anti-IL-23 antibodies have the sequences listed herein and are described in U.S. Patent No. 7,935,344, the entire contents of which are incorporated herein by reference.

[0042] Human antibodies specific for human IL-23 protein or fragments thereof can be raised against suitable immunogenic antigens, such as isolated IL-23 protein and / or portions thereof (including synthetic molecules such as synthetic peptides). Other specific or general mammalian antibodies can be similarly produced. Preparation of immunogenic antigens and production of monoclonal antibodies can be carried out using any suitable technique.

[0043] In one approach, a suitable immortal cell line (e.g., but not limited to, 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 heteromyelomas, fusion products thereof, or any cells or fusion cells derived therefrom, or any other suitable cell line known in the art) (see, for example, www.atcc.org, www.lifetech.com, etc.), antibody producing cells such as isolated or cloned spleen, peripheral blood, lymph, tonsil, or other immune or B cell containing cells, or recombinant or endogenous, viral, bacterial, or other nucleic acids, either as endogenous or heterologous. With any other cell expressing the heavy or light chain constant or variable, or framework or CDR sequences, either as algae, prokaryote, amphibian, insect, reptile, fish, mammal, rodent, horse, ovine, caprine, ovine, primate, eukaryote, genomic DNA, cDNA, rDNA, mitochondrial DNA or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single, double or triple stranded, hybridized, etc., or as any combination thereof, to produce a hybridoma. See, e.g., Ausubel, supra, and Colligan, Immunology, supra, Chapter 2, both of which are incorporated herein by reference in their entireties.

[0044] Antibody producing cells can also be obtained from the peripheral blood, or preferably the spleen or lymph nodes, of humans or other suitable animals immunized with the antigen of interest. Any other suitable host cells can also be used to express heterologous or endogenous nucleic acid encoding the antibody of the invention, specific fragments or variants thereof. Fused cells (hybridomas) or recombinant cells can be isolated using selective culture conditions or other suitable known methods and cloned by limiting dilution or cell sorting or other known methods. Cells producing antibodies with the desired specificity can be selected by a suitable assay (e.g., ELISA).

[0045] Other methods suitable for producing or isolating antibodies with the required specificity can be used, including, but not limited to, recombinant antibody selection from peptide or protein libraries (e.g., but not limited to, bacteriophage, ribosomal, oligonucleotide, RNA, cDNA, etc. display libraries, available, for example, from 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.).For example, European Patent No. 368,684, International Application No. GB91 / 01134, International Application No. GB92 / 01755, International Application No. GB92 / 002240, International Application No. GB92 / 00883, International Application No. GB93 / 00605, U.S. Patent Application No. 08 / 350260 (5 / 12 / 94), International Application No. GB94 / 01422, International Application No. GB94 / 02662, International Application No. Application No. GB97 / 01835, (CAT / MRC), WO 90 / 14443, WO 90 / 14424, WO 90 / 14430, International Application No. US94 / 1234, WO 92 / 18619, WO 96 / 07754, (Scripps), WO 96 / 13583, WO 97 / 08320 (MorphoSys), WO No. 95 / 16027 (BioInvent), WO 88 / 06630, WO 90 / 3809 (Dyax), U.S. Pat. No. 4,704,692 (Enzon), International Application No. US91 / 02989 (Affymax), WO 89 / 06283, European Patent No. 371998, European Patent No. 550400 (Xoma), European Patent No. 229046, No. 5,723,323, No. 5,763,192, No. 5,814,476, No. 5,817,483, No. 5,824,514, No. 5,976,862, WO 86 / 05803, EP 590689 (Ixsys), or stochastically generated peptides or proteins - see U.S. Pat. Nos. 5,723,323, 5,763,192, 5,814,476, 5,817,483, 5,824,514, 5,976,862, WO 86 / 05803, EP 590689 (Ixsys), Applied Molecular Evolution (Applied These techniques include those based on the National Institute of Molecular Evolution (AME), a predecessor of the American Association for Clinical Molecular Evolution (ACME), each of which is incorporated herein by reference in its entirety), or rely on the immunization of transgenic animals capable of producing a repertoire of human antibodies as known in the art and / or described herein (e.g., SCID mice, each of which is incorporated herein by reference in its entirety; Nguyen et al., Microbiol. Immunol. 41:901-907 (1997); Sandhu et al., Crit. Rev. Biotechnol. 16:95-118 (1996); Eren et al., Immunol. 93:154-161 (1998), and related patents and applications).Such techniques include ribosome display (Hanes et al., Proc. Natl. Acad. Sci. USA, 94:4937-4942 (May 1997); Hanes et al., Proc. Natl. Acad. Sci. USA, 95:14130-14135 (Nov. 1998)), single cell antibody production techniques (e.g., the selected 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 microdroplets and flow cytometry (Powell et al., J. Immunol. 1999, 11:111-111 (1996)). al., Biotechnol. 8:333-337 (1990); One Cell Systems, Cambridge, MA; Gray et al., J. Imm. Meth. 182:155-163 (1995); Kenny et al., Bio / Technol. 13:787-790 (1995)), B cell selectors (Steenbakkers et al., Molec. Biol. Reports 19:125-134 (1994); Jonak et al., Progress Biotech, Vol. 5, In Vitro Immunization in Hybridoma Technology, Borrebaeck, ed., Elsevier Science Publishers BV, Amsterdam, Netherlands (1988)).

[0046] Methods for engineering or humanizing non-human or human antibodies can also be used and are known in the art. Generally, a humanized or modified antibody has one or more amino acid residues derived from a non-human source, such as, but not limited to, mouse, rat, rabbit, non-human primate, or other mammalian source. These non-human amino acid residues are replaced by residues often referred to as "import" residues, which are typically taken from an "import" variable, constant, or other domain of a known human sequence.

[0047] Known human Ig sequences are, for example, :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 / immunology / 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, www.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.unam.mx / vir / V_mice.html, http: / / 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).

[0048] Such imported sequences can be used to reduce immunogenicity, or to reduce, enhance, or modify binding, affinity, binding rate constant, dissociation rate constant, avidity, specificity, half-life, or any other suitable property, as known in the art. Generally, CDR residues directly and most substantially affect antigen binding. Thus, non-human sequences in variable and constant regions can be replaced with human or other amino acids, while maintaining some or all of the non-human or human CDR sequences.

[0049] Antibodies may optionally be humanized, or human antibodies may be modified while retaining high affinity for the antigen and other favorable biological properties. To this end, humanized (or human) antibodies may optionally be prepared by a process of analysis of the parental sequences and various theoretical humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformations of selected candidate immunoglobulin sequences. Inspection of these displays allows analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., analysis of residues that influence the antigen-binding ability of the candidate immunoglobulin. In this way, framework (FR) residues can be selected and combined from the consensus and import sequences so that desired antibody properties, such as enhanced affinity for the target antigen, are achieved.

[0050] In addition, human IL-23 specific antibodies used in the methods of the invention may comprise a human germline light chain framework. In certain embodiments, the light chain germline sequence is selected from a human VK sequence, 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. In certain embodiments, the 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.

[0051] In other embodiments, the human IL-23 specific antibodies used in the methods of the invention may comprise a human germline heavy chain framework. In certain embodiments, the heavy chain human germline framework is selected from the group consisting of 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- 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.

[0052] In certain embodiments, the light chain variable region and / or the heavy chain variable region comprises a framework region, or at least a portion of a framework region (e.g., comprising 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 comprises a human consensus sequence for a particular framework (which are readily available at the sources of known human Ig sequences mentioned above). In other embodiments, at least FRH1, FRH2, FRH3, or FRH4 is a germline sequence (e.g., human germline) or comprises a human consensus sequence for a particular framework. In preferred embodiments, the framework regions are fully human framework regions.

[0053] Humanization or engineering of the antibodies of the present invention may be carried out in accordance with the methods described by Winter (Jones 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. USA 89:4285 (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. 5,76 6886, 5714352, 6204023, 6180370, 5693762, 5530101, 5585089, 5225539, 4816567, International Application No. US98 / 16280, US9 This can be done using any known method, such as, but not limited to, those described in US91 / 09630, US91 / 05939, US94 / 01234, International Application Nos. GB89 / 01334, GB91 / 01134, GB92 / 01755, International Publication Nos. WO 90 / 14443, WO 90 / 14424, WO 90 / 14430, and European Patent No. 229246 (each of which is incorporated by reference in its entirety, including the references cited therein).

[0054] In certain embodiments, the antibody comprises an altered (e.g., mutated) Fc region. For example, in some embodiments, the Fc region is altered 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 the amino acid modification with one or more further amino acid modifications that alter the C1q binding and / or complement dependent cytotoxicity function of the Fc region of the IL-23 binding molecule. Starting polypeptides of particular interest may be those that bind C1q and exhibit complement dependent cytotoxicity (CDC). Polypeptides with existing C1q binding activity, and optionally further with the ability to mediate CDC, may be modified to enhance one or both of these activities. Amino acid modifications that alter C1q and / or modify its complement dependent cytotoxicity function are described, for example, in WO 0042072, which is incorporated herein by reference.

[0055] As disclosed above, the Fc region of the human IL-23 specific antibody of the present invention can be engineered with altered effector functions, for example, by modifying C1q binding and / or FcγR binding, thereby altering complement-dependent cytotoxicity (CDC) activity and / or antibody-dependent cell-mediated cytotoxicity (ADCC) activity. An "effector function" serves to activate or reduce a biological activity (e.g., in a subject). Examples of effector functions include, but are not limited to, C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, downregulation of cell surface receptors (e.g., B cell receptor, BCR), and the like. Such effector functions may require the Fc region to bind to a binding domain (e.g., an antibody variable domain) and can be assessed using a variety of test methods (e.g., Fc binding assays, ADCC assays, CDC assays, and the like).

[0056] For example, variant Fc regions of human IL-23 (or anti-IL-23) antibodies can be generated that have improved C1q binding and improved FcγRIII binding (e.g., that have both improved ADCC activity and improved CDC activity). Alternatively, where it is desired to reduce or eliminate effector function, variant Fc regions can be modified to have reduced CDC activity and / or reduced ADCC activity. In other embodiments, only one of these activities may be enhanced, optionally with the other activity reduced at the same time (e.g., to generate Fc region variants with improved ADCC activity and reduced CDC activity, and vice versa).

[0057] Fc mutations can also be engineered and introduced to alter interactions with the neonatal Fc receptor (FcRn) and improve their pharmacokinetic properties. A 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).

[0058] Another type of amino acid substitution serves to modify the glycosylation pattern of the Fc region of a human IL-23 specific antibody. Glycosylation of the Fc region is typically either N-linked or O-linked. N-linked 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 sugars, N-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used. The recognition sequences for enzymatic attachment of a carbohydrate moiety to the asparagine side chain peptide sequence are asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline. Thus, the presence of either of these peptide sequences in a polypeptide provides a potential glycosylation site.

[0059] The glycosylation pattern can be modified, for example, by deleting one or more glycosylation sites found in the polypeptide and / or adding one or more glycosylation sites not present in the polypeptide. Addition of glycosylation sites to the Fc region of a human IL-23-specific antibody is conveniently accomplished by modifying the amino acid sequence to include one or more of the above tripeptide sequences (for N-linked glycosylation sites). A representative glycosylation variant has an amino acid substitution of residue Asn297 of the heavy chain. This modification may also be performed by the addition of, or substitution by, one or more serine or threonine residues to the original polypeptide sequence (for O-linked glycosylation sites). In addition, changing Asn 297 to Ala can remove one of the glycosylation sites.

[0060] In certain embodiments, the human IL-23 specific antibodies of the invention are expressed in cells expressing beta(1,4)-N-acetylglucosaminyltransferase III (GnT III), such that GnT III adds GlcNAc to the human IL-23 antibody. Methods for producing antibodies in such a manner are provided in WO 9954342, WO 03011878, JP 20030003097(A1), and Umana et al., Nature Biotechnology, 17:176-180, Feb. 1999, all of which are expressly incorporated herein by reference in their entirety.

[0061] Anti-IL-23 antibodies may also optionally be generated 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-23 antibodies may be isolated and immortalized from such animals using suitable methods, such as those described herein.

[0062] Transgenic mice capable of producing a repertoire of human antibodies that bind to human antigens can be produced using known methods (such as, but not limited to, U.S. Pat. Nos. 5,770,428, 5,569,825, 5,545,806, 5,625,126, 5,625,825, 5,633,425, 5,661,016, and 5,789,650 issued to Lonberg et al., WO 98 / 50433 to Jakobovits et al., WO 98 / 24893 to Jakobovits et al., WO 98 / 2491 to Lonberg et al., each of which is incorporated herein by reference in its entirety). No. 884 to Lonberg et al., International Publication No. WO 97 / 13852 to Lonberg et al., International Publication No. WO 94 / 25585 to Lonberg et al., International Publication No. WO 96 / 34096 to Kucherlapate et al., European Patent No. 0463151(B1) to Kucherlapate et al., European Patent No. 0710719(A1) to Kucherlapate et al., U.S. Pat. No. 5,545,807 to Surani et al., International Publication No. WO 90 / 04036 to Bruggemann et al., European Patent No. 0438474(B1) to Bruggemann et al., European Patent No. 0814259(A2) to Lonberg et al., UK Patent No. 2272440(A) to Lonberg et al., et al.Nature 368:856-859(1994),Taylor et al.,Int.Immunol.6(4)579-591(1994),Green et al,Nature Genetics 7:13-21(1994),Mendez et al.,Nature Genetics 15:146-156(1997),Taylor et al. 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)).Generally, these mice contain at least one transgene that comprises DNA derived from at least one human immunoglobulin locus that has been functionally rearranged, or is capable of undergoing functional rearrangement. The endogenous immunoglobulin loci of such mice can be disrupted or deleted to eliminate the ability of the mice to produce antibodies encoded by endogenous genes.

[0063] Screening of antibodies for specific binding to similar proteins or fragments can be successfully accomplished using peptide display libraries. The method involves screening a large collection of peptides for individual members with the desired function or structure. Antibody screening of peptide display libraries is well known in the art. The displayed peptide sequences can be 3-5000 or more amino acids long, frequently 5-100 amino acids long, and often about 8-25 amino acids long. In addition to direct chemical synthesis methods for creating peptide libraries, several recombinant DNA methods have also been described. One type involves the display of peptide sequences on the surface of bacteriophages or cells. Each bacteriophage or cell contains a nucleotide sequence that codes for a particular displayed peptide sequence. Such methods are described in WO 91 / 17271, WO 91 / 18980, WO 91 / 19818, and WO 93 / 08278.

[0064] Other systems for generating peptide libraries include aspects of both in vitro chemical synthesis and recombinant methods. See WO 92 / 05258, WO 92 / 14843, and WO 96 / 19256. See also U.S. Patent Nos. 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). See, e.g., U.S. Patent Nos. 4,704,692, 4,939,666, 4,946,778, 5,260,203, 5,455,030, 5,518,889, 5,534,621, 5,656,730, 5,763,733, 5,767,260, and 5,856,456 all assigned to Enzon; ​​U.S. Patent Nos. 5,223,409, 5,403,484, 5,571,698, and 5,837,500 all assigned to Dyax; U.S. Patent Nos. 5,427,908 and 5,580,717 all assigned to Affymax; Cambridge antibody See U.S. Patent No. 5,885,793 assigned to Eppendorf Technologies, U.S. Patent No. 5,750,373 assigned to Genentech, U.S. Patent Nos. 5,618,920, 5,595,898, 5,576,195, 5,698,435, 5,693,493, 5,698,417 assigned to Xoma, Colligan, supra, Ausubel, supra, or Sambrook, supra. Each of the above patents and publications is incorporated herein by reference in its entirety.

[0065] The antibodies used in the methods of the invention can also be prepared using nucleic acids encoding at least one anti-IL23 antibody to provide transgenic animals or mammals, such as goats, cows, horses, sheep, rabbits, etc., that produce such antibodies in their milk. Such animals can be provided using known methods. See, for example, but not limited to, U.S. Patent Nos. 5,827,690, 5,849,992, 4,873,316, 5,849,992, 5,994,616, 5,565,362, 5,304,489, etc., each of which is incorporated herein by reference in its entirety.

[0066] The antibodies used in the methods of the invention can further be prepared using at least one anti-IL23 antibody encoding nucleic acid to provide transgenic plants and cultured plant cells (e.g., but not limited to, tobacco and corn) that produce such antibodies, specific portions, or variants in plant parts or cells cultured therefrom. As a non-limiting example, transgenic tobacco leaves expressing recombinant proteins, for example, using an inducible promoter, have been successfully used to provide large quantities of recombinant proteins. See, e.g., Cramer et al., Curr. Top. Microbol. Immunol. 240:95-118 (1999) and references cited therein. Transgenic corn has also been used to express mammalian proteins at commercial production levels with biological activity equivalent to proteins produced in other recombinant systems or purified from natural sources. See, e.g., Hood et al., Adv. Exp. Med. Biol. 464:127-147 (1999) and references cited therein. Antibodies have also been produced in large quantities from transgenic plant seeds containing antibody fragments, such as single chain antibodies (scFv), including tobacco seeds and potato tubers. See, e.g., Conrad et al., Plant Mol. Biol. 38:101-109 (1998) and references cited therein. Thus, the antibodies of the present invention can also be produced using transgenic plants according to known methods. See, e.g., 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 references cited therein. Each of the above documents is incorporated herein by reference in its entirety.

[0067] The antibodies used in the methods of the present invention have a wide range of affinities (K D ) can bind human IL-23. In a preferred embodiment, the human mAb can optionally bind human IL-23 with high affinity. For example, the human mAb can bind human IL-23 at 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 -10 , 10 -11 , 10 -12 , 10 -13 K, or any range or value therein D can be combined with

[0068] The affinity or avidity of an antibody for an antigen can be determined experimentally using any suitable method. (See, e.g., Berzofsky, et al., "Antibody-Antigen Interactions," Fundamental Immunology, Paul, WE, Ed., Raven Press: New York, NY (1984); Kuby, Janis Immunology, WH Freeman and Company: New York, NY (1992), and methods described therein.) The affinity measured for a particular antibody-antigen interaction can be different when measured under different conditions (e.g., salt concentration, pH). Thus, affinity and other antigen binding parameters (e.g., K D , K a , K d Measurements of ) are preferably performed using standardized solutions of antibody and antigen, and standardized buffers, such as those described herein.

[0069] nucleic acid molecule Using the information provided herein, such as, for example, a nucleotide sequence encoding at least 70-100% of the contiguous amino acids of at least one of the light or heavy chain variable or CDR regions described herein, a particular fragment, variant, or consensus sequence thereof, or a deposited vector containing at least one of these sequences, among other sequences disclosed herein, a nucleic acid molecule of the invention encoding at least one anti-IL-23 antibody can be obtained using methods described herein or known in the art.

[0070] The nucleic acid molecules of the present invention may be in the form of RNA, such as mRNA, hnRNA, tRNA, or any other form, or in the form of DNA, including but not limited to cDNA and genomic DNA obtained by cloning or produced synthetically, or any combination thereof. The DNA may be triple-stranded, double-stranded, or single-stranded, or any combination thereof. Any portion of at least one strand of the DNA or RNA may be the coding strand, also known as the sense strand, or the non-coding strand, called the antisense strand.

[0071] The isolated nucleic acid molecules used in the methods of the present invention may include nucleic acid molecules that include an open reading frame (ORF) with at least one particular portion of at least one CDR, such as, but not limited to, at least one heavy or light chain CDR1, CDR2, and / or CDR3, optionally including one or more introns, nucleic acid molecules that include coding sequences for anti-IL-23 antibodies or variable regions, and nucleic acid molecules that include substantially different nucleotide sequences than those described above, but still encode at least one anti-IL-23 antibody described herein and / or known in the art due to the degeneracy of the genetic code. Of course, the genetic code is well known in the art. Thus, it would be routine for a person skilled in the art to generate such degenerate nucleic acid variants that encode the specific anti-IL-23 antibodies used in the methods of the present invention. See, for example, Ausubel et al., supra. Such nucleic acid variants are encompassed by the present invention. Non-limiting examples of isolated nucleic acid molecules include nucleic acids encoding HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3, respectively.

[0072] As described herein, nucleic acid molecules comprising nucleic acids encoding anti-IL-23 antibodies can include those that themselves encode the amino acid sequence of an antibody fragment, sequences encoding the full length of an antibody or a portion of an antibody, coding sequences for an antibody, fragment or portion, and additional sequences, such as at least one intron, with or without the aforementioned additional coding sequences, including, but not limited to, non-coding 5' and 3' sequences, e.g., transcribed non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals (e.g., ribosome binding and stability of mRNA), as well as additional non-coding sequences, but not limited to, coding sequences for at least one signal leader or fusion peptide, additional coding sequences encoding additional amino acids, e.g., amino acids that provide additional functions. Thus, the antibody coding sequence can be fused to a marker sequence, e.g., a marker sequence is a sequence that encodes a peptide that facilitates purification of an antibody comprising the antibody fragment or portion to which it is fused.

[0073] Polynucleotides that selectively hybridize to the polynucleotides described herein The method of the present invention uses isolated nucleic acids that hybridize under selective hybridization conditions to the polynucleotides disclosed herein. Thus, the polynucleotides of the present embodiment can be used to isolate, detect, and / or quantify nucleic acids that contain such polynucleotides. For example, the polynucleotides of the present invention can be used to identify, isolate, or amplify partial or full-length clones in a deposited library. In some embodiments, the polynucleotides are genomic or cDNA sequences that are isolated or otherwise complementary to cDNAs in a human or mammalian nucleic acid library.

[0074] Preferably, the cDNA library contains at least 80% of the full-length sequences, preferably at least 85% or 90% of the full-length sequences, more preferably at least 95% of the full-length sequences. The cDNA library can be normalized to increase the representation of rare sequences. Low or medium stringency hybridization conditions are typical, but not limited to, using sequences with low sequence identity to the complementary sequence. Optionally, medium and high stringency conditions can be used for sequences with higher identity. Low stringency conditions allow selective hybridization of sequences with about 70% sequence identity and can be used to identify orthologous or paralogous sequences.

[0075] Optionally, the polynucleotide encodes at least a portion of an antibody. The polynucleotide comprises a nucleic acid sequence that can be used for selective hybridization to a polynucleotide encoding an antibody of the present invention. See, e.g., Ausubel, supra; Colligan, supra, each of which is incorporated herein by reference in its entirety.

[0076] Nucleic acid construction An isolated nucleic acid can be produced using (a) recombinant methods, (b) synthetic techniques, (c) purification techniques, and / or (d) combinations thereof, as are well known in the art.

[0077] The nucleic acid may conveniently contain sequences in addition to the polynucleotide of the invention. For example, a multiple cloning site containing one or more endonuclease restriction sites may be inserted into the nucleic acid to aid in the isolation of the polynucleotide. Also, a translatable sequence may be inserted to aid in the isolation of the translated polynucleotide of the invention. For example, a hexahistidine marker sequence provides a convenient means for purifying the protein of the invention. The nucleic acid of the invention (excluding the coding sequence) is optionally a vector, adapter, or linker for cloning and / or expression of the polynucleotide of the invention.

[0078] Additional sequences can be added to such cloning and / or expression sequences to optimize their function in cloning and / or expression, to aid in the isolation of polynucleotides, or to improve the introduction of polynucleotides into cells. The use of cloning vectors, expression vectors, adapters, and linkers is well known in the art. (See, e.g., Ausubel, supra, or Sambrook, supra.)

[0079] Recombinant methods for constructing nucleic acids Isolated nucleic acid compositions, such as RNA, cDNA, genomic DNA, or any combination thereof, can be obtained from biological sources using any number of cloning methods known to those of skill in the art. In some embodiments, oligonucleotide probes that selectively hybridize under stringent conditions to the polynucleotides of the present invention are used to identify desired sequences in cDNA or genomic DNA libraries. Isolation of RNA and construction of cDNA and genomic libraries are well known to those of skill in the art. (See, e.g., Ausubel, supra, or Sambrook, supra).

[0080] Nucleic Acid Screening and Isolation Methods Probes based on the sequences of the polynucleotides used in the methods of the invention, such as those disclosed herein, can be used to screen cDNA or genomic libraries. Probes can be used to hybridize to genomic DNA or cDNA sequences to isolate homologous genes in the same or different organisms. Those skilled in the art will appreciate that various degrees of hybridization stringency can be used in the assay, and that either the hybridization or the wash medium can be made more stringent. The more stringent the hybridization conditions, the greater the degree of complementarity between the probe and the target at which duplex formation occurs. The degree of stringency can be controlled by one or more of temperature, ionic strength, pH, and the presence of a partially denaturing solvent, such as formamide. For example, the stringency of hybridization is successfully altered by changing the polarity of the reaction solution, for example, by manipulating the formamide concentration in the range of 0% to 50%. The degree of complementarity (sequence identity) required for detectable binding varies depending on the stringency of the hybridization medium and / or the wash medium. The degree of complementarity is optimally 100%, or 70-100%, or any range or value therein, however, it should be understood that minor differences in sequence in the probe and primers can be compensated for by reducing the stringency of the hybridization and / or wash medium.

[0081] Methods for amplifying RNA or DNA are well known in the art and can be used in accordance with the present invention without undue experimentation, based on the teachings and guidance provided herein.

[0082] Known methods of DNA or RNA amplification include the polymerase chain reaction (PCR) and related amplification processes (e.g., U.S. Pat. Nos. 4,683,195, 4,683,202, 4,800,159, and 4,965,188 to Mullis et al., U.S. Pat. Nos. 4,795,699 and 4,921,794 to Tabor et al., U.S. Pat. No. 5,142,033 to Innis, U.S. Pat. No. 5,122,464 to Wilson et al., U.S. Pat. No. 5,091,310 to Innis, U.S. Pat. No. 5,066,584 to Gyllensten et al., U.S. Pat. No. 5,066,584 to Gelfa et al., U.S. Pat. No. 5,122,464 to Innis, U.S. Pat. No. 5,091,310 ...122 No. 4,889,818 to nd et al., U.S. Patent No. 4,994,370 to Silver et al., U.S. Patent No. 4,766,067 to Biswas, and U.S. Patent No. 4,656,134 to Ringold), and RNA-mediated amplification (U.S. Patent No. 5,130,238 to Malek et al., trade name NASBA), which uses antisense RNA to a target sequence as a template for double-stranded DNA synthesis, the entire contents of which are incorporated herein by reference. (See, e.g., Ausubel, supra, or Sambrook, supra.)

[0083] For example, polymerase chain reaction (PCR) technology can be used to amplify the sequences of polynucleotides and related genes used in the methods of the present invention directly from genomic DNA or cDNA libraries. PCR and other in vitro amplification methods can also be useful, for example, for cloning nucleic acid sequences encoding proteins to be expressed, for generating nucleic acids to be used as probes for detecting the presence of desired mRNA in a sample, for sequencing nucleic acids, or for other purposes. Examples of techniques sufficient to guide the skilled artisan through in vitro amplification methods can be found in Berger, Sambrook, and Ausubel, supra, as well as U.S. Patent No. 4,683,202 to Mullis et al. (1987), and Innis, et al., PCR Protocols A Guide to Methods and Applications, Eds., Academic Press Inc., San Diego, CA (1990). Commercial kits for genomic PCR amplification are known in the art. See, for example, Advantage-GC Genomic PCR Kit (Clontech). In addition, for example, the T4 gene 32 protein (Boehringer Mannheim) can be used to improve yields of long PCR products.

[0084] Synthetic methods for constructing nucleic acids The isolated nucleic acid used in the method of the present invention can also be prepared by direct chemical synthesis by known methods (see, for example, Ausubel et al., supra). Chemical synthesis generally produces a single-stranded oligonucleotide that can be converted into double-stranded DNA by hybridization with a complementary sequence or by polymerization with a DNA polymerase using the single strand as a template. Those skilled in the art will recognize that chemical synthesis of DNA can be limited to sequences of about 100 or more bases, but longer sequences can be obtained by ligation of shorter sequences.

[0085] Recombinant Expression Cassettes The present invention employs recombinant expression cassettes comprising nucleic acids. Nucleic acid sequences, such as cDNA or genomic sequences encoding an antibody used in the methods of the present invention, can be used to construct recombinant expression cassettes that can be introduced into at least one desired host cell. Recombinant expression cassettes typically comprise a polynucleotide that is operably linked to a transcription initiation regulatory sequence that directs transcription of the polynucleotide in the intended host cell. Both heterologous and non-heterologous (i.e., endogenous) promoters can be utilized to direct expression of the nucleic acid.

[0086] In some embodiments, isolated nucleic acids that function as promoters, enhancers, or other elements can be introduced into a suitable location (upstream, downstream, or within an intron) of a non-heterologous form of a polynucleotide of the invention to up- or down-regulate expression of the polynucleotide. For example, endogenous promoters can be altered in vivo or in vitro by mutation, deletion, and / or substitution.

[0087] Vectors and host cells The present invention also relates to vectors comprising the isolated nucleic acid molecules, host cells engineered with the recombinant vectors, and the production of at least one anti-IL-23 antibody by recombinant techniques well known in the art (see, e.g., Sambrook et al., supra; Ausubel et al., supra, each of which is incorporated herein by reference in its entirety).

[0088] The polynucleotide can be optionally linked to a vector that contains a selection marker for propagation in the host.Generally, the plasmid vector is introduced into a precipitate such as a calcium phosphate precipitate or into a complex with a charged lipid.If the vector is a virus, it can be packaged in vitro using a suitable packaging cell line and then transduced into the host cell.

[0089] The DNA insert should be operably linked to a suitable promoter. The expression construct further comprises a transcription initiation 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 comprises translation beginning with a start and stop codon (e.g., UAA, UGA, or UAG) appropriately positioned at the end of the mRNA to be translated, with UAA and UAG being preferred for expression in mammalian or eukaryotic cells.

[0090] It is preferred, but optional, that the expression vector includes at least one selectable marker. Such markers include, for example, methotrexate (MTX), dihydrofolate reductase (DHFR, U.S. Pat. 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 for eukaryotic cell culture. synthetase, GS, U.S. Patent Nos. 5,122,464, 5,770,359, 5,827,739) resistance genes, and tetracycline or ampicillin resistance genes for culture in E. coli and other bacteria or prokaryotes (the above patents are incorporated herein by reference in their entireties). Appropriate culture media and conditions for the above host cells are known in the art. Suitable vectors will be readily apparent to those of ordinary skill in the art. Introduction of the vector construct into the host cell can be accomplished 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, such as Sambrook, supra, Chapters 1-4 and 16-18, and Ausubel, supra, Chapters 1, 9, 13, 15, 16.

[0091] At least one antibody used in the method of the invention may be expressed in modified form, such as a fusion protein, and may contain not only secretion signals, but also additional heterologous functional regions. For example, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the antibody to improve stability and persistence in the host cell during purification or during subsequent processing and storage. Peptide moieties may also be added to the antibody of the invention to facilitate purification. Such regions may be removed prior to final preparation of the antibody or at least one fragment thereof. Such methods are described in many standard laboratory manuals, such as Sambrook, supra, Chapters 17.29-17.42 and 18.1-18.74, Ausubel, supra, Chapters 16, 17, and 18.

[0092] Those skilled in the art are familiar with the many expression systems available for expressing the nucleic acid encoding the protein used in the method of the present invention. Alternatively, the nucleic acid can be expressed in a host cell by switching on (by manipulation) in the host cell containing the endogenous DNA encoding the antibody. Such methods are well known in the art, as described in U.S. Patent Nos. 5,580,734, 5,641,670, 5,733,746, and 5,733,761, which are incorporated herein by reference in their entirety.

[0093] An example of a cell culture useful for the production of an antibody, a particular portion or variant thereof is a mammalian cell. Mammalian cell lines are often in the form of a monolayer of cells, although suspensions or bioreactors of mammalian cells can also be used. A number of suitable host cell lines capable of expressing intact glycosylated proteins have been developed in the art, including COS-1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-1651), HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e.g., ATCC CRL1610) and BSC-1 (e.g., ATCC CRL-26) cell lines, Cos-7 cells, CHO cells, hep G2 cells, P3X63Ag8.653, SP2 / 0-Ag14, 293 cells, HeLa cells, and the like, which are readily available, for example, from the American Type Culture Collection (Manassas, Va.) (www.atcc.org). Preferred host cells include cells derived from lymphatic system, 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 cell is P3X63Ab8.653 or SP2 / 0-Ag14 cell.

[0094] Expression vectors for these cells can include one or more of the following expression control sequences, including, but not limited to, an origin of replication, a promoter (e.g., the late or early SV40 promoter, the CMV promoter (U.S. Pat. Nos. 5,168,062, 5,385,839), the HSV tk promoter, the pgk (phosphoglycerate kinase) promoter, the EF-1 alpha promoter (U.S. Pat. No. 5,266,491), at least one human immunoglobulin promoter, an enhancer, and / or processing information sites such as ribosome binding sites, RNA splice sites, polyadenylation sites (e.g., the SV40 large T Ag polyaddition site), and transcription termination sequences. See, e.g., Ausubel et al., supra; Sambrook et al., supra. Other cells useful for producing the nucleic acids or proteins of the invention are known and / or can be found, for example, in the American Type Culture Collection Catalogue of Cell Lines and These are available from Hybridomas (www.atcc.org) or other known or commercial sources.

[0095] When eukaryotic host cells are used, typically polyadenylation or transcription termination sequences are incorporated into the vector. An example of a termination sequence is the polyadenylation sequence from the bovine growth hormone gene. Sequences for accurate splicing of the transcript 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, gene sequences for controlling replication in host cells can be incorporated into the vector, as is known in the art.

[0096] Antibody purification Anti-IL-23 antibodies can be recovered and purified from recombinant cell cultures by well-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, hydroxylapatite chromatography, and lectin chromatography. High performance liquid chromatography ("HPLC") can also be used for purification. See, e.g., Colligan, Current Protocols in Immunology or Current Protocols in Protein Science, John Wiley & Sons, NY, NY (1997-2001), e.g., chapters 1, 4, 6, 8, 9, 10, each of which is incorporated herein by reference in its entirety.

[0097] Antibodies for use in the methods of the invention include naturally purified products, products of chemical synthetic processes, and products produced by recombinant techniques from eukaryotic hosts, including, for example, yeast, higher plants, insect, and mammalian cells. Depending on the host utilized in a recombinant production process, the antibody may be glycosylated or non-glycosylated, but is preferably glycosylated. Such methods are described in many standard laboratory manuals, such as Sambrook, supra, Sections 17.37-17.42; Ausubel, supra, Chapters 10, 12, 13, 16, 18, and 20; Colligan, Protein Science, supra, Chapters 12-14, all of which are incorporated herein by reference in their entirety.

[0098] Anti-IL-23 antibody. The anti-IL-23 antibodies of the present invention comprise at least a portion of an immunoglobulin molecule, such as, but not limited to, at least one ligand binding portion (LBP), such as, but not limited to, a heavy or light chain complementarity determining region (CDR) or a ligand binding portion thereof, a heavy or light chain variable region, a framework region (e.g., FR1, FR2, FR3, FR4, or a fragment thereof, and optionally, including at least one substitution, insertion, or deletion), a heavy or light chain constant region (e.g., at least one CDR, FR5, FR6, FR7, FR8, FR9, FR10, FR11, FR12, FR13, FR14, FR15, FR16, FR17, FR18, FR19, FR19, FR119, FR120, FR130, FR140, FR151, FR161, FR172, FR183, FR194, FR195, FR196, FR197, FR198, FR19 ... H 1, Hinge 1, Hinge 2, Hinge 3, Hinge 4, C H 2 or C H 3, or a fragment thereof, and optionally further comprising at least one substitution, insertion, or deletion), or any portion thereof. The antibody can include or be derived from any mammal, such as, but not limited to, human, mouse, rabbit, rat, rodent, primate, or any combination thereof.

[0099] The isolated antibodies used in the methods of the invention include the amino acid sequences of the antibodies disclosed herein encoded by any suitable polynucleotide, or any isolated or prepared antibody. Preferably, the human antibodies or antigen-binding fragments bind to human IL-23, thereby partially or substantially neutralizing at least one biological activity of the protein. Antibodies, or specified portions or variants thereof, that partially or preferably substantially neutralize at least one biological activity of at least one IL-23 protein or fragment, can bind to the protein or fragment, thereby inhibiting an activity mediated through binding of IL-23 to the IL-23 receptor, or through other IL-23-dependent or mediated mechanisms. As used herein, the term "neutralizing antibody" refers to an antibody that can inhibit 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, 100% or more, depending on the assay. The ability of an anti-IL-23 antibody to inhibit IL-23-dependent activity is preferably assessed by at least one suitable 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 a kappa or lambda light chain. In one embodiment, the human antibody comprises an IgG heavy chain or defined fragment, e.g., at least one of the following isotypes: IgG1, IgG2, IgG3, or IgG4 (e.g., γ1, γ2, γ3, γ4). Antibodies of this type can be prepared by utilizing transgenic mice or other non-human transgenic mammals that contain at least one human light chain (e.g., IgG, IgA, and IgM) transgene as 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.

[0100] The antibody binds to at least one particular epitope specific for at least one IL-23 protein, subunit, fragment, portion, or any combination thereof, which may comprise at least one antibody binding region that comprises at least a portion of the protein, preferably comprising at least one extracellular, soluble, hydrophilic, exoportion, or cytoplasmic portion of the protein.

[0101] Generally, a human antibody or antigen-binding fragment comprises an antigen-binding region that comprises at least one human complementarity determining region (CDR1, CDR2, and CDR3) or variant of at least one heavy chain variable region, and at least one human complementarity determining region (CDR1, CDR2, and CDR3) or variant of at least one light chain variable region. The CDR sequences can be derived from human germline sequences or can match exactly to germline sequences. For example, CDRs from a synthetic library derived from the original non-human CDRs can be used. These CDRs can be formed by incorporation of conservative substitutions from the original non-human sequence. In another specific embodiment, the antibody or antigen-binding portion or variant can have an antigen-binding region that comprises at least a portion of at least one light chain CDR (i.e., CDR1, CDR2, and / or CDR3) with the corresponding CDR1, 2, and / or 3 amino acid sequence.

[0102] Such antibodies can be prepared by preparing and expressing a nucleic acid molecule encoding the antibody (i.e., one or more) using conventional techniques involving recombinant DNA technology, or by chemically linking together the various portions of the antibody (e.g., CDRs, framework) using conventional techniques, or by using any other suitable method.

[0103] An anti-IL-23 specific antibody can comprise at least one of a heavy or light chain variable region having a defined amino acid sequence. For example, in a preferred embodiment, an anti-IL-23 antibody comprises at least one of a heavy chain variable region, optionally having the amino acid sequence of SEQ ID NO: 7, and / or at least one of a light chain variable region, optionally having the amino acid sequence of SEQ ID NO: 8. For example, in a preferred embodiment, an anti-IL-23 antibody comprises at least one of a heavy chain variable region, optionally having the amino acid sequence of SEQ ID NO: 9, and / or at least one of a light chain variable region, optionally having the amino acid sequence of SEQ ID NO: 10. Antibodies that bind to human IL-23 and comprise a defined heavy or light chain variable region can be prepared using suitable methods, such as those employing phage display (Katsube, Y., et al., Int J Mol. Med, 1(5):863-868 (1998)) or transgenic animals, as known in the art and / or described herein. For example, a transgenic mouse containing a functionally rearranged human immunoglobulin heavy chain transgene and a transgene containing DNA from a human immunoglobulin light chain locus capable of undergoing functional rearrangement can be immunized with human IL-23 or a fragment thereof to induce the production of antibodies. If desired, antibody-producing cells can be isolated and hybridomas or other immortalized antibody-producing cells can be prepared as described herein and / or known in the art. Alternatively, antibodies, particular portions or variants can be expressed using the encoding nucleic acid or a portion thereof in a suitable host cell.

[0104] The present invention also relates to antibodies, antigen-binding fragments, immunoglobulin chains and CDRs that contain 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 contain such chains or CDRs have high affinity (e.g., about 10 -9 K below M D) can bind to human IL-23. Amino acid sequences that are substantially the same as the sequences described herein include sequences containing conservative amino acid substitutions as well as amino acid deletions and / or insertions. A conservative amino acid substitution refers to the replacement of a first amino acid with a second amino acid that has chemical and / or physical properties (e.g., charge, structure, polarity, hydrophobicity / hydrophilicity) similar to those of the first amino acid. Conservative substitutions include, but are not limited to, replacing one amino acid with another 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.

[0105] Amino acid code The amino acids constituting the anti-IL-23 antibodies of the present invention are often abbreviated. Amino acid notation can be indicated by representing the amino acid by its one-letter code, its three-letter code, name, or three-nucleotide codon, and is well understood in the art (see Alberts, B., et al., Molecular Biology of The Cell, Third Ed., Garland Publishing, Inc., New York, 1994).

[0106] [Table 1]

[0107] The anti-IL-23 antibodies used in the methods of the invention may contain one or more amino acid substitutions, deletions, or additions, either by natural mutation or by human manipulation, as specified herein.

[0108] The number of amino acid substitutions that one of skill in the art may make will depend on many factors, including those described above. Generally, the number of amino acid substitutions, insertions, or deletions for any given anti-IL-23 antibody, fragment, or variant will not exceed 40, 30, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, e.g., 1-30, or any range or value therein, as specified herein.

[0109] Amino acids within an anti-IL-23 specific antibody that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (e.g., Ausubel, supra, Chapters 8, 15; Cunningham and Wells, Science 244:1081-1085 (1989)). The latter procedure introduces single alanine mutations at every residue in the molecule. The resulting mutant molecules are then tested for biological activity, including, but not limited to, at least one IL-23 neutralizing activity. Sites critical 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)).

[0110] An anti-IL-23 antibody can include, but is not limited to, at least one portion, sequence, or combination selected from all five of at least one of the contiguous amino acids of SEQ ID NOs: 1, 2, 3, 4, 5, and 6.

[0111] The IL-23 antibody or specific portion or variant may include, but is not limited to, at least one portion, sequence, or combination selected from at least 3 to 5 contiguous amino acids of the above SEQ ID NO:, 5 to 17 contiguous amino acids of the above SEQ ID NO:, 5 to 10 contiguous amino acids of the above SEQ ID NO:, 5 to 11 contiguous amino acids of the above SEQ ID NO:, 5 to 7 contiguous amino acids of the above SEQ ID NO:, or 5 to 9 contiguous amino acids of the above SEQ ID NO:.

[0112] The anti-IL-23 antibody can further optionally comprise at least one polypeptide of 70-100% of 5, 17, 10, 11, 7, 9, 119, or 108 contiguous amino acids of SEQ ID NO: above. In one embodiment, the amino acid sequence of an immunoglobulin chain or portion thereof (e.g., variable region, CDR) has about 70-100% identity (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 therein) with the amino acid sequence of the corresponding chain of at least one of the SEQ ID NO: above. For example, the amino acid sequence of the light chain variable region can be compared to the sequence of the SEQ ID NO: above, or the amino acid sequence of the heavy chain CDR3 can be compared to the sequence of the SEQ ID NO: above. 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 known in the art.

[0113] "Identity," as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as determined by the match between strings of such sequences. "Identity" and "similarity" can be readily calculated by known methods, including, but 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). Additionally, percent identity values ​​can be obtained from amino acid and nucleotide sequence alignments generated using default settings in AlignX, a component of Vector NTI Suite 8.0 (Informax, Frederick, MD).

[0114] Preferred methods for determining identity are designed to obtain the highest degree of correspondence between the sequences 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)). BLAST X programs are publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBINLM NIH Bethesda, Md. 20894: Altschul, S., et al., J. Mol. Biol. 215:403-410 (1990). The well-known Smith Waterman algorithm may also be used to determine identity.

[0115] Preferred parameters for polypeptide sequence comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison matrix: BLOSSUM62 from Hentikoff and Hentikoff, Proc. Natl. Acad. Sci, USA. 89: 10915-10919 (1992), Gap penalty: 12 Gap length penalty: 4 A program useful with these parameters is publicly available as the "Gap" program from the Genetics Computer Group, Madison Wis. The aforementioned parameters are the default parameters for peptide sequence comparisons (as well as no penalty for end gaps).

[0116] Preferred parameters for polynucleotide comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48:443-453 (1970) Comparison matrix: match = +10, mismatch = 0 Gap penalty: 50 Gap length penalty: 3 Available as the "Gap" program from the Genetics Computer Group, Madison Wis. These are the default parameters for nucleic acid sequence comparisons.

[0117] By way of example, a polynucleotide sequence may be identical to another sequence, i.e., 100% identical, or may contain up to a certain integer number of nucleotide alterations compared to a reference sequence. Such alterations may be selected from the group consisting of deletion, substitution (including transitions and transversions), or insertion of at least one nucleotide, which may occur at the 5' or 3' terminal position of the reference nucleotide sequence, or anywhere between these terminal positions, and may be distributed either individually among the nucleotides of the reference sequence, or in one or more adjacent groups within the reference sequence. The number of nucleotide alterations may be determined by multiplying the total number of nucleotides in the sequence by the numerical percentage of the corresponding percent identity (divided by 100) and subtracting the product from the total number of nucleotides in the sequence, or is determined by n.sub.n.ltorsim.x.sub.n-(x.sub.ny), where n.sub.n is the number of nucleotide alterations, x.sub.n is the total number of nucleotides in the sequence, and y is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95%, etc., and any non-integer product of x.sub.n and y is rounded down to the nearest integer before subtraction from x.sub.n.

[0118] Modification of a polynucleotide sequence encoding the above SEQ ID NO may create nonsense, missense, or frameshift mutations in the coding sequence, thereby modifying the polypeptide encoded by the polynucleotide after such modification. Similarly, a polypeptide sequence may be identical to the reference sequence of the above SEQ ID NO, i.e., 100% identical, or may contain up to a certain integer number of amino acid modifications compared to the reference sequence such that the percent identity is less than 100%. Such modifications are selected from the group consisting of deletion, substitution (including conservative and non-conservative substitution), or insertion of at least one amino acid, which may occur at the amino or carboxy terminal position of the reference polypeptide sequence or anywhere between these terminal positions, and may be distributed either individually between the amino acids of the reference sequence, or in one or more adjacent groups within the reference sequence. The number of amino acid changes for a given percent identity may be calculated by multiplying the total number of amino acids in the above SEQ ID NO by the numerical percent of the respective percent identity (divided by 100) and subtracting the product from the total number of amino acids in the above SEQ ID NO, or is determined by n.sub.a.ltorsim.x.sub.a-(x.sub.ay), where n.sub.a is the number of amino acid changes, x.sub.a is the total number of amino acids in the sequence ID, and y is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, etc., and any non-integer product of x.sub.a and y is rounded down to the nearest integer before subtraction from x.sub.a.

[0119] Exemplary heavy and light chain variable region sequences, and portions thereof, are set forth in the SEQ ID NOs: above. An antibody of the invention, or a specified variant thereof, can include any number of contiguous amino acid residues from an antibody of the invention, the number being selected from the group of integers consisting of 10-100% of the number of contiguous residues in an anti-IL-23 antibody. Optionally, this subsequence of contiguous amino acids is 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 amino acids in length, or any range or value therein. Additionally, the number of such subsequences can be any integer selected from the group consisting of 1-20, such as at least 2, 3, 4, or 5.

[0120] As will be appreciated by those of skill in the art, the present invention includes at least one biologically active antibody of the invention. A biologically active antibody has a specific activity that is at least 20%, 30%, or 40%, preferably at least 50%, 60%, or 70%, and most preferably at least 80%, 90%, or 95% to 100% or more (including but not limited to up to 10-fold the specific activity) of that of a natural (non-synthetic), endogenous, or related and known antibody. Methods for assaying and quantitatively measuring enzymatic activity and substrate specificity are well known to those of skill in the art.

[0121] In another aspect, the invention relates to human antibodies and antigen-binding fragments described herein that are modified by the covalent attachment of an organic moiety. Such modifications can produce antibodies or antigen-binding fragments with improved pharmacokinetic properties (e.g., increased serum half-life in vivo). The organic moiety can 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 has a molecular weight of about 800 to about 120,000 daltons and can be a polyalkane glycol (e.g., polyethylene glycol (PEG), polypropylene glycol (PPG)), a carbohydrate polymer, an amino acid polymer, or polyvinylpyrrolidone, and the fatty acid group or fatty acid ester group can contain about 8 to about 40 carbon atoms.

[0122] The modified antibodies and antigen-binding fragments can include one or more organic moieties that are directly or indirectly covalently attached to the antibody. Each organic moiety attached to the antibody or antigen-binding fragment of the present invention can be independently a hydrophilic polymer group, a fatty acid group, or a fatty acid ester group. As used herein, the term "fatty acid" includes monocarboxylic and dicarboxylic acids. As used herein, the term "hydrophilic polymer group" refers to an organic polymer that is more soluble in water than octane. For example, polylysine is more soluble in water than octane. Thus, antibodies modified by the covalent attachment of polylysine are encompassed by the present invention. Hydrophilic polymers suitable for modifying the antibodies of the present invention can 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 that modify the antibodies of the present invention have, as individual molecular entities, a molecular weight of about 800 to about 150,000 daltons. For example, PEG 5000 and PEG20,000 can be used, where the subscript is the average molecular weight of the polymer in Daltons. The hydrophilic polymer group can be substituted with 1 to about 6 alkyl groups, fatty acid groups, or fatty acid ester groups. Hydrophilic polymers substituted with fatty acid or fatty acid ester groups can be prepared by utilizing a suitable method. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer.

[0123] Fatty acids and fatty acid esters suitable for modifying antibodies of the invention may be saturated or may contain one or more units of unsaturation. Fatty acids suitable for modifying antibodies of the invention include, for example, n-dodecanoate (C 12 , laurate), n-tetradecanoate (C 14 , myristate), n-octadecanoate (C 18 , stearate), n-eicosanoate (C 20 , arachidate), n-docosanoate (C 22 , behenic acid), n-triacontanoate (C 30 ), n-tetracontanoate (C 40 ), cis-Δ9-octadecanoate (C 18 oleate), all cis-Δ5,8,11,14-eicosatetraenoate (C 20 , arachidonate), octanedioic acid, tetradecanedioic acid, octadecanedioic acid, docosanedioic acid, and the like. Suitable fatty acid esters include monoesters of dicarboxylic acids containing a straight or branched chain lower alkyl group. The lower alkyl group can contain from 1 to about 12, preferably from 1 to about 6, carbon atoms.

[0124] Modified human antibodies and antigen-binding fragments can be prepared using suitable methods, such as by reacting with one or more modifying agents. As used herein, the term "modifying agent" refers to a suitable organic group (e.g., hydrophilic polymer, fatty acid, fatty acid ester) that contains an activating group. An "activating group" is a chemical moiety or functional group that can react with a second chemical group under appropriate conditions, thereby forming a covalent bond between the modifying agent and the second chemical group. For example, amine-reactive activating groups include electrophilic groups such as tosylate, mesylate, halo (chloro, bromo, fluoro, iodo), N-hydroxysuccinimidyl ester (NHS), and the like. Activating groups that can react with thiols include, for example, maleimide, iodoacetyl, acrylolyl, pyridyl disulfide, 5-thiol-2-nitrobenzoic acid thiol (TNB-thiol), and the like. 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. Suitable 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 attached directly to organic groups (e.g., hydrophilic polymers, fatty acids, fatty acid esters) or to linker moieties, such as divalent C1-C 12The linker moiety may be linked via a group (wherein one or more carbon atoms may be replaced by a heteroatom 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-. A modifying agent containing a linker moiety may be produced, for example, by reacting a mono-Boc-alkyldiamine (e.g., mono-Boc-ethylenediamine, mono-Boc-diaminohexane) with a fatty acid in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) to form 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 coupled to another carboxylate as described, or it can be reacted with maleic anhydride and the resulting product cyclized to produce an activated maleimide derivative of a fatty acid (see, e.g., WO 92 / 16221 (Thompson et al.), the entire teachings of which are incorporated herein by reference).

[0125] Modified antibodies can be produced by reacting a human antibody or antigen-binding fragment with a modifying agent. For example, an organic moiety can be attached to the antibody in a non-site-specific manner using an amine-reactive modifying agent, e.g., an NHS ester of PEG. Modified human antibodies or antigen-binding fragments can also be prepared by reducing disulfide bonds (e.g., intrachain disulfide bonds) of an antibody or antigen-binding fragment. The reduced antibody or antigen-binding fragment can then be reacted with a thiol-reactive modifying agent to produce the modified antibody of the invention. Modified human antibodies and antigen-binding fragments containing organic moieties attached to specific sites of the antibodies of the invention can be prepared using suitable 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).

[0126] The methods of the invention also employ anti-IL-23 antibody compositions comprising at least one, at least two, at least three, at least four, at least five, at least six, or more of said anti-IL-23 antibodies provided in a non-naturally occurring composition, mixture, or form as described herein and / or known in the art. Such compositions include non-naturally occurring compositions comprising at least one or two full-length, C- and / or N-terminal deletion mutants, domains, fragments, or specific variants of an anti-IL-23 antibody amino acid sequence selected from the group consisting of 70-100% of the contiguous amino acids of the SEQ ID NOs. above, or specific fragments, domains, or variants thereof. A preferred anti-IL-23 antibody composition comprises at least one or two full-length, fragments, domains, or variants as at least one CDR- or LBP-containing portion of an anti-IL-23 antibody sequence described herein, e.g., 70-100% of the SEQ ID NOs. above, or specific fragments, domains, or variants thereof. More preferred compositions comprise, for example, 70-100% of at least one of the above SEQ ID NOs, or specific fragments, domains, or variants thereof, 40-99%. Such composition percentages may be by weight, volume, concentration, molarity, or molar concentration as a liquid or dry solution, mixture, suspension, emulsion, particle, powder, or colloid, as known in the art or as described herein.

[0127] Antibody compositions containing further therapeutically active ingredients The antibody compositions used in the methods of the invention can optionally further comprise an effective amount of at least one compound or protein selected from at least one of anti-infective agents, cardiovascular (CV) system acting agents, central nervous system (CNS) agents, autonomic nervous system (ANS) agents, respiratory agents, gastrointestinal (GI) tract acting agents, hormonal agents, fluid or electrolyte balancing agents, hemodynamic agents, anti-neoplastic agents, immunomodulatory agents, ophthalmic, otic or nasal agents, topical agents, nutritional agents, etc. Such drugs are well known in the art, including the formulations, indications, dosages, and administration of each as set forth herein (see, e.g., Nursing 2001 Handbook of Drugs, 21st Edition, each of which is incorporated herein by reference in its entirety). st edition, Springhouse Corp., Springhouse, PA, 2001; Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ; Pharmacotherapy Handbook, Wells et al., Appleton & Lange, Stamford, CT).

[0128] As examples of drugs that can be combined with the antibodies of the method of the present invention, the anti-infective drug can be at least one selected from amebicide or at least one antiprotozoal drug, anthelmintic drug, antifungal drug, antimalarial drug, antituberculous drug or at least one antilepromycotic drug, aminoglycoside, penicillin, cephalosporin, tetracycline, sulfonamide, fluoroquinolone, antiviral drug, macrolide anti-infective drug, and miscellaneous anti-infective drugs. The hormonal drug can be at least one selected from corticosteroid, androgen, or at least one anabolic steroid, estrogen, or at least one progestin, gonadotropin, antidiabetic drug, or at least one glucagon, thyroid hormone, thyroid hormone antagonist, pituitary hormone, and parathyroid mimetic drug. The at least one cephalosporin can be at least one selected from cefaclor, cefadroxil, cefazolin sodium, cefdinir, cefepime hydrochloride, cefixime, cefmetazole sodium, cefonicid sodium, cefoperazone sodium, cefotaxime sodium, cefotetan disodium, cefoxitin sodium, cefpodoxime proxetil, cefprozil, ceftazidime, ceftibuten, ceftizoxime sodium, ceftriaxone sodium, cefuroxime axetil, cefuroxime sodium, cephalexin hydrochloride, cephalexin monohydrate, cephradine, and loracarbef.

[0129] The at least one corticosteroid can be at least one selected from betamethasone, betamethasone acetate or betamethasone sodium phosphate, betamethasone sodium phosphate, cortisone acetate, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, fludrocortisone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone cypionate, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, methylprednisolone, methylprednisolone acetate, methylprednisolone sodium succinate, prednisolone, prednisolone acetate, prednisolone sodium phosphate, prednisolone tebutate, prednisone, triamcinolone, triamcinolone acetonide, and triamcinolone diacetate. The at least one androgenic or anabolic steroid can be at least one selected from danazol, fluoxymesterone, methyltestosterone, nandrolone decanoate, nandrolone phenpropionate, testosterone, testosterone cypionate, testosterone enanthate, testosterone propionate, and a testosterone transdermal system.

[0130] The at least one immunosuppressant can be at least one selected from azathioprine, basiliximab, cyclosporine, daclizumab, lymphocyte immunoglobulin, muromonab-CD3, mycophenolate mofetil, mycophenolate mofetil hydrochloride, sirolimus, and tacrolimus.

[0131] The at least one topical anti-infective can be at least one 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, naftifine hydrochloride, neomycin sulfate, nitrofurazone, nystatin, silver sulfadiazine, terbinafine hydrochloride, terconazole, tetracycline hydrochloride, tioconazole, and tolnaftate. The at least one scabicide or pediculicide can be at least one selected from crotamiton, lindane, permethrin, and pyrethrins. The at least one topical corticosteroid can be at least one selected from betamethasone dipropionate, betamethasone valerate, clobetasol propionate, desonide, desoximetasone, dexamethasone, dexamethasone sodium phosphate, diflorasone diacetate, fluocinolone acetonide, fluocinonide, flurandrenolide, fluticasone propionate, halcionide, hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone valerate, mometasone furoate, and triamcinolone acetonide. (See, e.g., pages 1098-1136 of Nursing 2001 Drug Handbook.)

[0132] Anti-IL-23 antibody compositions comprise at least one anti-IL-12 / 23p40 or IL-23 antibody that is contacted with or administered to a cell, tissue, organ, animal, or subject in need of such modulation, treatment, or therapy, and optionally at least one TNF antagonist (such as, but not limited to, a TNF chemical or protein antagonist, a TNF monoclonal or polyclonal antibody or fragment, a soluble TNF receptor (e.g., p55, p70, or p85) or fragment, a fusion polypeptide thereof, or a small molecule TNF antagonist, such as TNF binding protein I or II (TBP-1 or TBP-II), nerelimonmab, infliximab, or ribozyme inhibitors (e.g., ribozyme inhibitors), ... The composition or pharmaceutical composition may further include at least one of any suitable and effective amount of compositions or pharmaceutical compositions further including at least one selected from the group consisting of antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, etanercept, gold sodium thiomalate, 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-40, etc. (e.g., IL-1, IL-2, etc.). Suitable dosages are well known in the art. For example, Wells et al., eds., Pharmacotherapy Handbook, 2 nd Edition, Appleton and Lange, Stamford, CT (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, CA (2000), each of which is incorporated herein by reference in its entirety.

[0133] The anti-IL-23 antibody compound, composition, or mixture used in the method of the present invention may further comprise at least one of any suitable auxiliary agent, such as, but not limited to, a diluent, a binder, a stabilizer, a buffer, a salt, a lipophilic solvent, a preservative, an adjuvant, etc. Pharmaceutically acceptable auxiliary agents are preferred. Methods for preparing such sterile solutions and non-limiting examples thereof are well known in the art and are described, for example, in Gennaro, Ed., Remington's Pharmaceutical Sciences, 1896, 1999, 1998, 19 ... th Edition, Mack Publishing Co. (Easton, PA) 1990. Pharmaceutically acceptable carriers suitable for the mode of administration, solubility, and / or stability of the anti-IL-23 antibody, fragment, or variant compositions well known in the art or described herein can be routinely selected.

[0134] Pharmaceutical excipients and additives useful in the present compositions include, but are not limited to, proteins, peptides, amino acids, lipids, and carbohydrates (e.g., saccharides including monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides, derivatized sugars such as alditols, aldonic acids, esterified sugars, and polysaccharides or sugar polymers), which may be present alone or in combination and comprise 1-99.99% by weight or volume, alone or in combination. Representative protein excipients include serum albumins such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acids / antibody components that may also function in a buffering capacity include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. One preferred amino acid is glycine.

[0135] Carbohydrate excipients suitable for use in the present invention include, for example, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, etc., disaccharides such as lactose, sucrose, trehalose, cellobiose, etc., polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starches, etc., alditols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), myo-inositol, etc. Preferred carbohydrate additives for use in the present invention are mannitol, trehalose, and raffinose.

[0136] The anti-IL-23 antibody composition may also include a buffer or pH adjusting agent, typically a salt prepared from an organic acid or base. Representative buffers include organic acid salts such as salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid, Tris, tromethamine hydrochloride, or phosphate buffers. A preferred buffer for use in the composition is an organic acid salt such as citric acid.

[0137] Additionally, anti-IL-23 antibody compositions may include polymeric excipients / additives such as polyvinylpyrrolidone, Ficoll (a polymeric sugar), dextrates (e.g., cyclodextrins such as 2-hydroxypropyl-β-cyclodextrin), polyethylene glycol, flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatic agents, surfactants (e.g., polysorbates such as "TWEEN 20" and "TWEEN 80"), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0138] These and additional known pharmaceutical additives and / or excipients suitable for use in the anti-IL-23 antibody, portion or variant compositions according to the invention are known in the art and are described, for example, in "Remington: The Science & Practice of Pharmacy," 1999. thed., Williams&Williams, (1995) and “Physician's Desk Reference”, 52 nd ed., Medical Economics, Montvale, NJ (1998), the disclosures of which are incorporated herein by reference in their entireties. Preferred carrier or excipient materials are carbohydrates (e.g., monosaccharides and alditols) and buffers (e.g., citric acid) or polymeric agents. Exemplary carrier molecules are mucopolysaccharides, hyaluronic acid, which may be useful for intra-articular delivery.

[0139] formulation As mentioned above, the present invention provides stable formulations, preferably saline or phosphate buffer with selected salts, as well as preservative-containing preservative solutions and formulations, and versatile preservative formulations suitable for pharmaceutical or veterinary use, comprising at least one anti-IL-23 antibody in a pharma- ceutically acceptable formulation. The preservative formulations include at least one known preservative, optionally selected from the group consisting of at least one phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrate, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate), alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof, in an aqueous diluent. As known in the art, the range may be from 0.001 to 5%, or any range or value therein, 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, 1.7, 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, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 3.1, 3.2, 3.3, 3.4, 3.5, 3.7, 3.8, 3.9, 3.1, 3.2 ...3, 3.4, 3.5, 3.8, 3.9, 3.1, 3.2, 3.3, 3.4, 3.5, Any suitable concentration or mixture may be used, such as 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 therein.Non-limiting examples include no preservatives, 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 of suitable alkylparabens include 0.05, 0.25, 0.28, 0.5, 0.9, 1.0%), 0.0005 to 1.0% alkylparaben (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, 1.0%), and the like.

[0140] As discussed above, the methods of the invention employ articles of manufacture comprising packaging and at least one vial containing a solution of at least one anti-IL-23 specific antibody, optionally with a buffer and / or preservative formulated in an aqueous diluent, the packaging comprising a label indicating that such solution can be maintained for 1, 2, 3, 4, 5, 6, 9, 12, 18, 20, 24, 30, 36, 40, 48, 54, 60, 66, 72 hours or more. The invention further employs articles of manufacture comprising packaging and a first vial containing a lyophilized anti-IL-23 specific antibody and a second vial containing an aqueous diluent of the formulated buffer or preservative, the packaging comprising a label instructing a patient to reconstitute the anti-IL-23 specific antibody with the aqueous diluent to form a solution that can be maintained for 24 hours or more.

[0141] The anti-IL-23 specific antibodies used in accordance with the present invention may be produced by recombinant means, including production from mammalian cells or transgenic preparations, or may be purified from other biological sources, as described herein or known in the art.

[0142] Ranges of anti-IL-23 specific antibodies include those amounts that will give a concentration of about 1.0 μg / mL to about 1000 mg / mL upon reconstitution for wet / dry systems, although lower and higher concentrations are workable and depend on the intended delivery vehicle, e.g., for solution formulations, as opposed to transdermal patch, pulmonary, transmucosal, or osmotic or micropump methods.

[0143] Preferably, the aqueous diluent further comprises a pharma- ceutically acceptable preservative, optionally. Preferred preservatives include those selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof. The concentration of the preservative used in the formulation is sufficient to produce an antimicrobial effect. Such concentration will vary depending on the preservative selected, and can be easily determined by those skilled in the art.

[0144] Other excipients, such as isotonicity agents, buffers, antioxidants, and preservative enhancers, can be optionally and preferably added to the diluent. An isotonicity agent, such as glycerin, is generally used at a known concentration. A physiologically tolerable buffer is preferably added to provide improved pH control. The formulations can cover a wide range of pH, such as from about pH 4 to about pH 10, and preferably from about pH 5 to about pH 9, and most preferably from about 6.0 to about 8.0. 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, especially phosphate buffered saline (PBS).

[0145] Other additives, such as pharma- ceutically acceptable solubilizers, such as Tween 20 (polyoxyethylene (20) sorbitan monolaurate), Tween 40 (polyoxyethylene (20) sorbitan monopalmitate), Tween 80 (polyoxyethylene (20) sorbitan monooleate), Pluronic F68 (polyoxyethylene polyoxypropylene block copolymer), and PEG (polyethylene glycol), or non-ionic 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 pumps or plastic containers are used to administer the formulation. The presence of pharma-ceutically acceptable surfactants reduces the tendency of proteins to aggregate.

[0146] The formulations can be prepared by a process that includes mixing at least one anti-IL-23 antibody with a preservative selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof, in an aqueous diluent. Mixing at least one anti-IL-23 specific antibody with a preservative in an aqueous diluent is performed using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a quantity of at least one anti-IL-23 specific antibody in a buffer solution is combined with a desired preservative in a sufficient amount of buffer solution to provide the desired concentration of protein and preservative. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of the components, the use or non-use of additional additives, the temperature and pH during preparation of the formulation are all factors that can be optimized for the administration concentration and administration means used.

[0147] The formulation can be provided to the patient as a clear solution or as a dual vial containing a vial of lyophilized anti-IL-23 specific antibody reconstituted with a second vial containing water, preservatives and / or excipients, preferably phosphate buffer and / or saline, and the selected salt in an aqueous diluent. Either the single solution vial or the dual vial requiring reconstitution can be reused multiple times to satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0148] The product is useful for administration over a period ranging from immediate to 24 hours or more. Thus, the products claimed by the present invention provide significant benefits to patients. The formulations of the present invention can optionally be safely stored at temperatures between about 2° C. and about 40° C. and retain the biological activity of the protein for extended periods of time, and thus the packaging label can state that the solution can be stored and / or used for 6, 12, 18, 24, 36, 48, 72, or 96 hours or more. When using a preserved diluent, such label can include use up to 1-12 months, half a year, one and a half years, and / or up to two years.

[0149] A solution of anti-IL-23 specific antibody can be prepared by a process that includes 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 in an amount sufficient to provide the desired concentration of protein, and optionally a preservative or buffer. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of the components, whether or not additional additives are used, the temperature and pH at which the formulation is prepared are all factors that can be optimized for the administration concentration and administration means used.

[0150] The claimed products can be provided to patients as clear solutions or as dual vials containing at least one lyophilized vial of an anti-IL-23 specific antibody that is reconstituted with a second vial containing an aqueous diluent. Either the single solution vial or the dual vial requiring reconstitution can be reused multiple times to satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0151] The claimed products can be provided indirectly to patients by providing a pharmacy, clinic, or other such institution or facility with a dual vial containing a clear solution or a vial of at least one lyophilized anti-IL-23 specific antibody reconstituted with a second vial containing an aqueous diluent, where the clear solution can be up to a liter or even more in volume, from which smaller amounts of the at least one antibody solution can be withdrawn one or more times from the larger container and transferred to smaller vials and provided to customers and / or patients by the pharmacy or clinic.

[0152] Approved devices that include single vial systems include pen-type injection 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®, and the like (Becton Dickensen (Franklin Suitable devices include those manufactured or developed by National Medical Products, Weston Medical (Peterborough, UK, www.weston-medical.com), Medi-Ject Corp (Minneapolis, MN, www.mediject.com), and similar suitable devices. Recognized devices that include dual vial systems include pen-type syringe systems, such as HumatroPen®, for reconstituting lyophilized medication in a cartridge to deliver the reconstituted solution. Examples of suitable other devices include pre-filled syringes, auto-injectors, needleless syringes, and needleless IV infusion sets.

[0153] The product may include packaging. The packaging provides the conditions under which the product may be used, as well as any information required by regulatory agencies. The packaging of the present invention, when applicable, provides instructions to the patient to reconstitute at least one anti-IL-23 antibody with an aqueous diluent to form a solution and use the solution for a period of 2-24 hours or more in a wet / dry two vial product. In the case of a single vial solution product, pre-filled syringe, or autoinjector, the label indicates that such solution may be used for a period of 2-24 hours or more. The product is useful for human pharmaceutical product applications.

[0154] The formulations used in the methods of the present invention can be prepared by a process that includes mixing an anti-IL-23 antibody and a selected buffer, preferably saline or a phosphate buffer containing a selected salt. Mixing the anti-IL-23 antibody and the buffer in an aqueous diluent is performed using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a quantity of at least one antibody in water or buffer is combined with a 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 the components, whether or not additional additives are used, the temperature and pH at which the formulation is prepared are all factors that can be optimized for the administration concentration and administration means used.

[0155] The method of the present invention provides pharmaceutical compositions including various formulations that are useful and acceptable for administration to human or animal patients. Such pharmaceutical compositions are prepared using "standard" water as a diluent and routine methods well known to those skilled in the art. For example, buffer components such as histidine and histidine monohydrochloride hydrate may be provided first, followed by the addition of an appropriate non-final volume of "standard" water diluent, sucrose, and polysorbate 80. The isolated antibody may then be added. Finally, the volume of the pharmaceutical composition is adjusted to the desired final volume under "standard" conditions using water as a diluent. Those skilled in the art will recognize several other methods suitable for the preparation of pharmaceutical compositions.

[0156] A pharmaceutical composition may be an aqueous solution or suspension containing the indicated mass of each component per volume unit of water or having the indicated pH at "standard conditions". As used herein, the term "standard conditions" refers to a temperature of 25°C ± 2°C and a pressure of 1 atmosphere. The term "standard conditions" is not used in the art to refer to a single set of art-recognized temperatures or pressures, but is instead a reference condition that specifies the 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 temperature and pressure. Those skilled in the art will recognize that pharmaceutical compositions equivalent to those disclosed herein can be produced at other temperatures and pressures. The equivalence of such pharmaceutical compositions to those disclosed herein should be determined under "standard conditions" conditions defined above (e.g., 25°C ± 2°C and a pressure of 1 atmosphere).

[0157] Importantly, such a pharmaceutical composition may contain "about" a certain value of the mass of a component (e.g., "about 0.53 mg of L-histidine") or have a certain value of the pH value per unit volume of the pharmaceutical composition. The mass or pH value of a component present in the pharmaceutical composition is "about" a given numerical value when the isolated antibody is present in the pharmaceutical composition or when the isolated antibody is present in the pharmaceutical composition after it is removed from the pharmaceutical composition (e.g., by dilution) and can bind to a peptide chain. That is, the mass value or pH value or other value of a component is "about" a given numerical value when the binding activity of the isolated antibody is maintained and detectable after the isolated antibody is placed in the pharmaceutical composition.

[0158] Competitive binding analysis is performed to determine whether IL-23 specific mAbs bind similar or different epitopes and / or compete with each other. Abs are individually coated onto ELISA plates. Competing mAbs are added, followed by biotinylated hrIL-23. For a positive control, the same mAb for coating can be used as a competing mAb ("self-competition"). IL-23 binding is detected using streptavidin. These results indicate whether the mAbs recognize similar or partially overlapping epitopes on IL-23.

[0159] One aspect of the methods of the invention involves administering a pharmaceutical composition to a patient.

[0160] In one embodiment of the pharmaceutical composition, the isolated antibody concentration is about 77 to about 104 mg per mL of pharmaceutical composition. In another embodiment of the pharmaceutical composition, the pH is about 5.5 to about 6.5.

[0161] The stable or preserved formulations can be provided to patients as clear solutions or as dual vials containing a vial of at least one lyophilized anti-IL-23 antibody that is reconstituted with a second vial containing a preservative or buffer and additives in an aqueous diluent. Either the single solution vial or the dual vial requiring reconstitution can be reused multiple times to satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0162] Other formulations or methods of stabilizing anti-IL-23 antibodies may be other than clear solutions of lyophilized powders containing the antibodies. Non-clear solutions include formulations containing particulate suspensions, which are compositions containing anti-IL-23 antibodies in structures of various sizes known variously as microspheres, microparticles, nanoparticles, nanospheres, or liposomes. Such relatively homogeneous essentially spherical particulate formulations containing active agents can be formed by contacting an aqueous phase containing active agents and polymers with a non-aqueous phase, as taught in U.S. Pat. No. 4,589,330, and then evaporating the non-aqueous phase to cause coalescence of particles from the aqueous phase. Porous microparticles can be prepared by using a first phase containing active agents and polymers dispersed in a continuous solvent, and removing the solvent from the suspension by lyophilization or dilution-extraction-precipitation, as taught in U.S. Pat. No. 4,818,542. Preferred polymers for such preparations are gelatin agar, starch, arabinogalactan, albumin, collagen, polyglycolic acid, polylactic acid, glycolide-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 acids), poly(2-hydroxyethyl DL-aspartamide), poly(ester urea), poly(L-phenylalanine / ethylene glycol), poly( ... The preferred polymers are natural or synthetic copolymers or polymers selected from the group consisting of poly(methyl methacrylate), poly(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). Solvents useful for dissolving the polymer and / or active agent include water, hexafluoroisopropanol, methylene chloride, tetrahydrofuran, hexane, benzene, or hexafluoroacetone sesquihydrate.The process of dispersing the active-containing phase into the second phase can include forcing the first phase through orifices in a nozzle under pressure to affect droplet formation.

[0163] Dry powder formulations may also result from processes other than freeze-drying, such as, for example, spray drying, or solvent extraction by evaporation, or by precipitation of a crystalline composition followed by one or more steps to remove the aqueous or non-aqueous solvent. The preparation of spray-dried antibody formulations is taught in U.S. Pat. No. 6,019,968. Antibody-based dry powder compositions can be produced by spray drying a solution or slurry of antibody and, optionally, excipients in a solvent under conditions to provide a respirable dry powder. Solvents include polar compounds, such as water and ethanol, that are easily dried. 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 that typically includes a hydrofluoroalkane propellant, as taught in WO 9916419. The stabilized dispersion can be administered to the lungs of a patient using a metered dose inhaler. Equipment useful in the commercial production of spray dried drugs is manufactured by Buchi Ltd. or Niro Corp.

[0164] Anti-IL-23 antibodies, either in the stable or preserved formulations or in solution described herein, can be administered to a patient in accordance with the present invention via a variety of delivery methods well known in the art, such as SC or IM injection, transdermal, transpulmonary, transmucosal, implants, osmotic pumps, cartridges, micropumps, or other means understood by those of skill in the art.

[0165] Therapeutic Applications The present invention also provides methods for modulating or treating SSc in a cell, tissue, organ, animal, or patient using at least one IL-23 antibody of the invention known in the art or described herein, e.g., by administering to or contacting the cell, tissue, organ, animal, or patient with a therapeutically effective amount of an IL-23 specific antibody.

[0166] Any of the methods of the invention may comprise administering to a cell, tissue, organ, animal, or patient in need of such modulation, treatment, or therapy an effective amount of a composition or pharmaceutical composition comprising an anti-IL-23 antibody. Such methods may optionally further comprise co-administration or combination therapy for the treatment of such a disease or disorder, wherein administering the at least one anti-IL-23 antibody, specified portion, or variant thereof, is in combination with at least one TNF antagonist (such as, but not limited to, a chemical or proteinaceous TNF antagonist, a TNF monoclonal or polyclonal antibody or fragment, a soluble TNF receptor (e.g., p55, p70, or p85) or fragment, a fusion polypeptide thereof, or a small molecule TNF antagonist, such as a TNF binding protein. TBP-1 or TBP-II, nerelimonumab, infliximab, etanercept (Enbrel™), adalimumab (Humira™), CDP-571, CDP-870, afelimomab, lenercept, etc.), antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, sodium aurothiomalate, hydroxychloroquine sulfate, leflunomide, sulfasalazine), muscle relaxants, narcotics, non-steroidal anti-inflammatory drugsdrugs, NSAIDs), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blockers, antibacterials (e.g., aminoglycosides, antifungals, antiparasitics, antivirals, carbapenams, cephalosporins, fluoroquinolones, macrolides, penicillins, sulfonamides, tetracyclines, other antibacterials), psoriasis medications, corticosteroids, anabolic steroids, diabetes medications, minerals, nutritional drugs, thyroid medications, vitamins, calcium-related hormones, antidiarrheals, antitussives, antiemetics, antineoplastics, laxatives, anticoagulants, erythropoietin (e.g., epoetin alfa), filgrastim (e.g., G-CSF, Neupogen), sargramostim (GM-CSF, Leukine), The method further comprises administering, before, simultaneously, and / or after at least one selected from an immunological agent, an immunoglobulin, an immunosuppressant (e.g., basiliximab, cyclosporine, daclizumab), a growth hormone, a hormone replacement drug, an estrogen receptor modulator, a mydriatic, a cycloplegic, an alkylating agent, an antimetabolite, a mitotic inhibitor, a radiopharmaceutical, an antidepressant, an antimanic drug, an antipsychotic, an anxiolytic, a hypnotic, a sympathomimetic, a stimulant, donepezil, tacrine, an asthma drug, a beta agonist, an inhaled steroid, a leukotriene inhibitor, a methylxanthine, a cromolyn, an epinephrine or an analogue, dornase alfa (Pulmozyme), a cytokine, or a cytokine antagonist. Suitable dosages are well known in the art. See, for example, Wells et al., eds., Pharmacotherapy Handbook, 2 nd 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, 21 stedition, Springhouse Corp., Springhouse, PA, 2001; Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ, each of which is incorporated herein by reference in its entirety.

[0167] Treatment Typically, treatment of SSc is effected by administering an effective amount or dose of an anti-IL-23 antibody composition, which, depending on the specific activity of the active agent contained in the composition, ranges from at least about 0.01 to 500 milligrams of anti-IL-23 antibody per kilogram of patient, on average, in total, per dose, and preferably at least about 0.1 to 100 milligrams of antibody per kilogram of patient, per single or multiple doses. Alternatively, an effective serum concentration may include a serum concentration of 0.1 to 5000 μg / mL, per single or multiple doses. Suitable dosages are known to medical practitioners and will, of course, depend on the specific disease state, the specific activity of the composition being administered, and the specific patient being treated. In some cases, it may be necessary to provide repeated doses, i.e., repeated individual doses of a particular monitored or metered dose, to achieve the desired therapeutic dose, where the individual doses are repeated until the desired daily dose or effect is obtained.

[0168] Preferred doses are optionally 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70 , 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 and / or 100-500 mg / kg / dose, or any range, value, or fraction thereof, or 0.1, 0.5, 0.9, 1.0, 1.1, 1.2, 1.5, 1.9, 2.0, 2.5, 2.9, 3.0, 3.5, 3.9, 4.0, 4.5, 4.9, 5.0, 5.5, 5.9, 6.0, 6.5, 6.9, 7.0, 7.0, 7.5 ... .5, 7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 20, 12.5, 12.9, 13.0, 13.5, 13.9, 14.0, 14.5, 4.9, 5.0, 5.5., 5.9, 6.0, 6.5, 6.9 , 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 12, 12.5, 12.9, 13.0, 13.5, 13.9, 14, 14.5, 15, 15.5, 15.9, 16, 16.5, 16.9 , 17, 17.5, 17.9, 18, 18.5, 18.9, 19, 19.5, 19.9, 20, 20.5, 20.9, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 96, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, and / or 5000 μg / mL, or any range, value or fraction thereof.

[0169] Alternatively, the dose administered may vary depending on known factors such as the pharmacodynamic characteristics of the particular agent and its method and route of administration, the age, health and weight of the recipient, the nature and extent of the symptoms, type of concurrent treatment, frequency of treatment, and the desired effect. The dose of active ingredient may typically be about 0.1 to 100 milligrams per kilogram of body weight. Typically, 0.1 to 50, preferably 0.1 to 10 milligrams per kilogram per dose, or in sustained release form, is effective to obtain the desired results.

[0170] As non-limiting examples, treatment of humans or animals may be performed using a single dose, intravenous administration, or multiple doses over 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 days. or additionally, on at least one day of eyes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 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, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 109, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 11 0.1 to 100 mg / kg per day, for example, 0.5, 0.9, 1.0, 1.2, 1.4, 1.6, 1.8, 1.9, 1.10, 1.20, 1.40, 1.65, 1.80, 1.95, 1.99, 1.99, 1.06, 1.07, 1.08, 1.09, 1.10, 1.11, 1.12, 1.13, 14, 15, 16, 17, 18, 19, or 20 years, or any combination thereof. The antibody may be provided as a single or periodic dose of 0, 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90, or 100 mg / kg of at least one antibody of the invention.

[0171] Dosage forms (compositions) suitable for internal administration generally contain about 0.001 milligrams to about 500 milligrams of active ingredient per unit or container. In these pharmaceutical compositions, the active ingredient is usually present in an amount of about 0.5 to 99.999% by weight based on the total weight of the composition.

[0172] For parenteral administration, the antibody may be formulated as a solution, suspension, emulsion, particle, powder, or lyophilized powder, provided together with or separately from a pharma- ceutical acceptable parenteral vehicle. Examples of such vehicles are water, saline, Ringer's solution, dextrose solution, and 1-10% human serum albumin. Non-aqueous vehicles such as liposomes and fixed oils can also be used. The vehicle or lyophilized powder may contain additives that maintain isotonicity and chemical stability (e.g., sodium chloride for isotonicity, mannitol, buffers and preservatives for chemical stability). The formulation is sterilized by known or suitable techniques.

[0173] Suitable pharmaceutical carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, A. Osol, a standard reference text in this field.

[0174] Alternative Administration Many known and developed modes can be used in accordance with the present invention to administer a pharma- ceutical effective amount of an anti-IL-23 antibody. Although pulmonary administration is used in the following description, other modes of administration may be used in accordance with the present invention with suitable results. The IL-23 specific antibodies of the present invention can be delivered in a carrier, as a solution, emulsion, colloid or suspension, or as a dry powder, using any of a variety of devices and methods suitable for administration by inhalation or by other methods described herein or known in the art.

[0175] Parenteral Formulation and Administration Preparations for parenteral administration may contain sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, hydrogenated naphthalenes, etc. as common excipients. Aqueous or oily suspensions for injection can be prepared by using appropriate emulsifiers or wetting agents and suspending agents according to known methods. Injections can be non-toxic parenterally administrable diluents such as aqueous solutions, sterile injections, or suspensions in solvents. Usable vehicles or solvents include water, Ringer's solution, isotonic saline, etc., and sterile fixed oils can be used as normal solvents or suspension solvents. For these purposes, any kind of fixed oils and fatty acids, including natural, synthetic, or semi-synthetic fatty oils or fatty acids, 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 means, the gas pressurized needleless injection device described in U.S. Pat. No. 5,851,198, and the laser perforator device described in U.S. Pat. No. 5,839,446, which are incorporated herein by reference in their entireties.

[0176] alternative delivery The invention further relates to administration of anti-IL-23 antibodies by parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intrabronchial, intraabdominal, intracapsular, intrachondral, intrasinus, intracavity, intracerebellar, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, intralesional, bolus, intravaginal, rectal, buccal, sublingual, intranasal or transdermal means. The anti-IL-23 antibody compositions can be formulated for use parenterally (subcutaneously, intramuscularly, or intravenously) or for any other administration, particularly in the form of liquid solutions or suspensions, particularly in semi-solid forms such as, but not limited to, creams and suppositories, for use in vaginal or rectal administration, in forms such as, but not limited to, tablets or capsules, for buccal or sublingual administration, or intranasally, in forms such as, but not limited to, powders, nasal drops or aerosols, or certain agents, or with chemical enhancers such as dimethylsulfoxide to either modify the skin structure or increase the drug concentration in transdermal patches (Junginger, et al. In "Drug Permeation Enhancement;" Hsieh, DS, Eds., pp. 59-90 (Marcel Dekker, Inc. New York, 1999), which are incorporated herein by reference in their entirety). 1994), or application of protein and peptide containing formulations to the skin (WO 98 / 53847), or application of an electric field to create a transient transport pathway, such as electroporation, or to increase the mobility of a charged drug through the skin, such as iontophoresis, or application of ultrasound, such as sonophoresis (U.S. Pat. Nos. 4,309,989 and 4,767,402), for transdermal use, such as, but not limited to, gels, ointments, lotions, suspensions, or patch delivery systems (the above publications and patents are incorporated herein by reference in their entireties).

[0177] Having generally described the present invention, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended to be limiting. Further details of the present invention are illustrated by the following non-limiting examples. The disclosures of all citations herein are expressly incorporated herein by reference.

[0178] Example 1 - Treatment of systemic sclerosis with IL23 antibodies A Multicenter, Randomized, Placebo-Controlled, Double-Blind, Proof-of-Concept Study of Guselkumab in Participants With Systemic Sclerosis Guselkumab (CNTO1959) is a fully human immunoglobulin G1 lambda (IgG1λ) monoclonal antibody (mAb) that binds to human interleukin (IL)-23 with high specificity and affinity. Guselkumab binds to the p19 subunit of IL-23, thereby blocking the binding of cell surface IL-23 receptors to extracellular IL-23, inhibiting IL-23-specific intracellular signaling and subsequent cytokine production. In this way, guselkumab inhibits the biological activity of IL-23 in all in vitro assays examined.

[0179] [Table 2]

[0180] [Table 3]

[0181] hypothesis In-house analysis of ribonucleic acid (RNA) sequencing (RNA-seq) data obtained from skin samples from patients with recent-onset SSc showed upregulation of IL-23 / IL17-related genes (analysis from the global Translational Science Medicine team).

[0182] This is a proof-of-concept (PoC) study aimed to detect early efficacy signals of guselkumab compared to placebo in participants with SSc to support further clinical development. ● The null hypothesis is that the treatment difference between guselkumab and placebo for the primary endpoint is equal to zero. ● The alternative hypothesis is that the treatment difference between guselkumab and placebo for the primary endpoint is not zero.

[0183] Due to the PoC nature of this study, a two-sided alpha of 0.2 is assigned to test this PoC hypothesis. The PoC objective is considered to be met if the p-value calculated to test this PoC hypothesis is less than 0.2.

[0184] Clinical Endpoint Description Modified Rodnan Skin Score The mRSS is a validated physical examination method to estimate skin hardening. It correlates with biopsies measuring skin thickness and reflects prognosis and visceral complications, especially in early disease. It is scored on an ordinal scale from 0 (normal) to 3 (severe hardening) across 17 body regions, with a maximum score of 51, and is used to classify the severity of SSc. It is widely used as a primary and secondary outcome measure in randomized clinical trials. The assessment should be performed by a physician with experience and training in skin scoring. To prevent interobserver variability, the same physician must perform skin scoring on the same participants throughout the study.

[0185] American College of Rheumatology Composite Response Index for Diffuse Cutaneous Systemic Sclerosis Score The ACR CRISS is a composite response index for clinical trials in early dcSSc developed by an international group of SSc experts. Application of the ACR CRISS algorithm in randomized clinical trials is a two-step process. First, participants are assessed whether they meet the criteria for "not improved." If yes, these participants are assigned a probability score of 0.0.

[0186] The investigator will assess whether the participant meets the criteria: New scleroderma renal crisis A decline in predicted FVC% of 15% or more (relative) should be considered with another FVC test within 1 month, an HRCT to confirm ILD (if a previous chest HRCT did not show ILD), and a predicted * those confirmed by FVC less than 80% of the value. New onset of left ventricular failure (defined as left ventricular ejection fraction 45% or less) requiring treatment * New onset of PAH during right-sided cardiac catheterization requiring treatment * * Caused by systemic sclerosis

[0187] For the remaining participants, a probability will be calculated based on change in five measures: mRSS, % of predicted FVC, HAQ-DI, Patient Global Assessment, and Physician Global Assessment, where each measure has a probability score between 0 and 1. The ACR CRIS score will be calculated by the sponsor.

[0188] Pulmonary function tests The following pulmonary function measurements are performed locally: Percentage of FVC and predicted FVC - DLCO (mL / min / mmHg) and percent of predicted DLCO (corrected for hemoglobin). Oxygen saturation data is also collected.

[0189] Doctor's overall evaluation The subjects' PhGA-assessed global SSc status will be assessed using a 10 cm visual analogue scale, with a score of 0 = excellent and a score of 10 = very poor.

[0190] Health Assessment Questionnaire-Disability Index The Stanford HAQ is a brief self-report questionnaire that assesses physical function for activities of daily living across eight domains: dressing and grooming, sitting, eating, walking, hygiene, reach, grip, and activity. Although it was initially developed for use in RA, it has been successfully applied to a variety of rheumatic conditions, including idiopathic inflammatory myopathies. The Stanford HAQ translated into Japanese with culturally appropriate modifications to questions in the sitting, eating, and reach categories will be used in the study.

[0191] Frequency scale of symptoms of gastroesophageal reflux disease (FSSG) The FSSG questionnaire is a brief questionnaire for assessing esophageal symptoms of GERD and is validated based on endoscopic esophagitis. The original version of the FSSG consists of the seven most common acid reflux-related symptoms of GERD and five motility disorder-related symptoms, with higher scores indicating more underlying GERD. Each score was determined as follows: 0=none, 1=occasionally, 2=sometimes, 3=frequently, and 4=always.

[0192] PRO devices will be provided in local languages ​​in accordance with local guidelines. ● The PRO device must be available for regulatory and Institutional Review Board (IRB) / Independent Ethics Committee (IEC) submissions and therefore the PRO device or screenshots should be provided in a companion manual accompanying the protocol or with the device submitted with the protocol. ● PRO and AE data are not adjusted to each other.

[0193] Digital ulcer evaluation Digital ulcers are defined as full thickness (greater than 3 mm in diameter) skin lesions with loss of epithelium, including crusted lesions (note that minor pitting scars and hyperkeratotic lesions are excluded). Healing is defined by re-epithelialization with absence of pain and exudate. Digital ulcer assessments will be performed by a designated investigator during the study. Ulcer counts and ulcer burden will be calculated by the sponsor.

[0194] Nail fold capillaroscope Nailfold capillaroscopy is a non-invasive method to visualize nailfold capillaries and evaluate microvascular morphology. Nailfold capillary abnormalities are included in the ACR / EULAR classification criteria for SSc. Nailfold capillary abnormalities are evaluated by using nailfold capillaroscopy. Typical changes in nailfold capillaries during SSc include total number of capillaries, capillary dimensions, capillary morphology, hemorrhage, blood flow velocity in the capillaries, and nailfold length. These changes are calculated. Visual medium is photographed.

[0195] High-resolution computed tomography (HRCT) evaluation Pulmonary complications are one of the most important causes of morbidity and mortality in patients with SSc. With recent advances in computer-based scoring, analysis of CT images has attracted attention as a method to evaluate fibrotic changes in ILD. Texture-based analysis and computer vision-based approaches can be applied to imaging data and used to evaluate the degree of pulmonary fibrosis in chest CT images. This evaluation should be performed centrally.

[0196] Biomarkers In the optional biomarker substudy, biopsies of non-lesional (unaffected) skin samples at week 0 and lesional (week 0) skin samples at weeks 0 and 24 will be obtained from all consenting participants.

[0197] Data collected from these samples will be used for exploratory testing, including but not limited to the following purposes: • Understand the molecular effects of guselkumab. • Understand the etiology of SSc. • Understand why individual participants may respond differently to guselkumab. • Understand the impact of guselkumab treatment on skin or systemic inflammation. • Develop a diagnostic test to identify SSc or SSc populations that may be responsive or non-responsive to treatment with guselkumab.

[0198] Overall Plan This is a randomized, double-blind, placebo-controlled, parallel, multicenter intervention study to evaluate the efficacy of guselkumab in men and women aged 18 to 75 years (inclusive), with SSc diagnosed according to the American College of Rheumatology (ACR) and European League Against Rheumatism (EULAR) 2013 criteria, disease duration (defined as the time from the first non-Raynaud's phenomenon episode) ≤36 months, and mRSS ≥10 and ≤22.

[0199] Participants will be randomly assigned in a 1:1 ratio to one of the following treatment arms based on randomization strata of presence of ILD (yes, no), baseline mRSS (low [≥10, ≤15] or high [≥16, ≤22]), and baseline anti-topoisomerase I antibody status (positive, negative): ● Guselkumab (group / arm A): Guselkumab 400 mg intravenously (IV) was administered at weeks 0, 4, and 8 (induction), followed by guselkumab 200 mg subcutaneously (SC) every 4 weeks (Q4W) from weeks 12 to 48 (maintenance). ● Placebo (Group / Arm B): Matching placebo was administered IV at weeks 0, 4, and 8 (induction), followed by matching placebo administered SC Q4W from weeks 12 to 48 (maintenance).

[0200] The main study will be conducted in three phases: a screening phase of up to 6 weeks, a double-blind intervention phase of 52 weeks, and a safety follow-up phase with a post-intervention follow-up visit 12 weeks after the participant's last study medication intervention to collect any AEs since the last study visit.

[0201] Participants who complete the main study (Weeks 0 to 52; i.e., after the Week 48 assessment and before the Week 52 assessment) and who, in the investigator's opinion, could benefit from continued treatment, will participate in the LTE by signing an ICF before or at Week 52. Individual participation duration will be approximately 66 weeks without LTE and 118 weeks with LTE.

[0202] Three planned database locks (DBLs) will occur: when all participants have completed the week 24 visit (hereafter referred to as the "week 24 DBL"), the end of study (EOS) of the main study (week 52 visit for participants who joined the LTE or week 60 visit for participants who did not join the LTE, hereafter referred to as the "week 52 DBL"), and the EOS of the LTE (week 112 visit, hereafter referred to as the "final DBL"). Note that if the LTE is terminated according to the Internal Decision Committee (IDC) recommendation (section λ.5), there will be only two DBLs (week 24 and week 52 DBLs) and LTE data collected for participants who have already joined the LTE at the time of the week 24 DBL will be analyzed in the week 52 DBL along with data up to the week 52 or week 60 visit.

[0203] Key efficacy assessments include mRSS, ACR CRISS score, pulmonary function tests: FVC and percentage of predicted FVC, DLCO (mL / min / mmHg) and percentage of predicted DLCO (corrected for hemoglobin), PhGA, and PGA. Key safety assessments include monitoring for AEs (including SAEs, AEs of special interest [AESIs], infections, injection site reactions, adverse events related to infusion transiently, and hypersensitivity reactions), physical examinations, vital sign measurements, electrocardiography (ECG) measurements, laboratory tests, ILD (assessed by centrally read HRCT), and tuberculosis (TB) assessments. Additionally, PK, biomarker, and immunogenicity assessments will also be performed in this study.

[0204] Number of participants The trial will target to enroll approximately 56 participants in total, with 28 participants planned per intervention arm.

[0205] Intervention group and duration Participants will be randomly assigned in a 1:1 ratio to one of the following study intervention arms: ● Guselkumab (group / arm A): Guselkumab 400 mg IV at weeks 0, 4, and 8 (induction), followed by guselkumab 200 mg SC Q4W from weeks 12 to 48 (maintenance). ● Placebo (Group / Arm B): Matching placebo administered IV at weeks 0, 4, and 8 (induction), followed by matching placebo administered SC Q4W from weeks 12 to 48 (maintenance).

[0206] During the LTE, all participants will receive guselkumab treatment in the LTE as follows, depending on the arm they are in during the main study: ● Group / Arm A (guselkumab arm from the main study): Guselkumab 200 mg SC and placebo IV at weeks 52, 56, and 60 of the LTE, followed by guselkumab 200 mg SC Q4W from weeks 64 to 100 of the LTE. ● Group / Arm B (placebo arm from the main study): Placebo administered SC and guselkumab 400 mg IV at weeks 52, 56, and 60 of LTE, followed by guselkumab 200 mg SC Q4W from weeks 64 to 100 of LTE.

[0207] Individual participation periods were approximately 66 weeks without LTE and 118 weeks with LTE.

[0208] Efficacy evaluation Efficacy assessment includes the following: ● mRSS ACR CRISS score Pulmonary function tests (PFTs): - FVC and percentage of predicted FVC - DLCO (mL / min / mmHg) and percentage of predicted DLCO (corrected for hemoglobin). ● PhGA Patient-reported outcomes (PROs): - P.G.A. - HAQ-DI - FSSG Questionnaire ● Digital ulcer assessment Nailfold capillaroscopy ● Lung HRCT (read centrally)

[0209] Pharmacokinetic evaluation Serum samples will be analyzed to determine guselkumab concentrations using respective validated, specific and sensitive methods by, or under the supervision of, the sponsor's respective assay.

[0210] Immunogenicity assessment Serum samples are screened for antibodies that bind to guselkumab, and titers of confirmed positive samples are reported. Serum samples positive for anti-guselkumab antibodies are further characterized for neutralizing antibodies (Nab) to guselkumab. Other analyses may be performed to verify the stability of anti-guselkumab antibodies and / or to further characterize the immunogenicity of guselkumab.

[0211] autoantibodies At screening, samples are collected to assess the presence of autoantibodies (including, but not limited to, anti-ribonucleic acid [RNA] polymerase, anti-centromere, and anti-topoisomerase).

[0212] Pharmacodynamics and Biomarkers Peripheral blood mononuclear cells (PBMC) and serum samples will be collected at the time points specified in the evaluation schedule (SoA) to evaluate blood cell and molecular biomarkers and allow assessment of target engagement in tissue / efficacy PD readouts.

[0213] In the optional biomarker substudy, biopsies of non-lesional (unaffected) skin samples at week 0 and lesional (week 0) skin samples at weeks 0 and 24 will be obtained from all consenting participants.

[0214] Pharmacogenetic evaluation Pharmacogenetic blood samples will be collected, if necessary, from participants who consent individually to this component of the study to allow for pharmacogenetic investigations. Participants' participation in pharmacogenetic studies is voluntary.

[0215] Safety assessment Safety assessments performed at each study visit include evaluation of AEs (occurring at the visit and between evaluation visits), ILD monitoring (assessed by centrally read HRCT), TB and other infection assessments, clinical laboratory blood tests (hematology and serum chemistry including C-reactive protein), pregnancy testing, physical examination, ECG measurements, vital signs, concomitant medication review, and observation for injection site reactions, hypersensitivity reactions, and AEs temporally related to the infusion. Additionally, for participants who are eligible for surface antigen (HBsAg) negative, core antibody (anti-HBc) and / or surface antibody (anti-HBs) positive and have a negative HBV DNA test, HBV DNA quantification should be monitored at least at 3-month or shorter intervals.

[0216] statistical methods Sample size determination A 95% confidence interval (CI) of [-4.7,-1.7] in the change in mRSS from baseline to 6 months is considered a clinically meaningful change. The lower limit of this 95% CI value, -4.7, has been adopted as the expected treatment difference in the sample size calculation for this study, assuming that guselkumab will be able to achieve clinically meaningful changes in most guselkumab-treated participants.

[0217] Assuming a standard deviation (SD) of 8 points, 80% power at a two-sided significance level of 0.20, and a randomization ratio of 1:1, a total of at least 54 participants would be required to detect a treatment effect of -4.7 points. Allowing for up to 2 randomized participants, but with no post-baseline efficacy assessments, we plan for approximately 56 randomized participants (28 each).

[0218] Population for the analysis set Analyses of efficacy and participant information will include all randomized participants who received at least one dose (full or partial) of the study intervention and will be analyzed based on the randomized treatment group, regardless of the treatment they actually received.

[0219] The safety analysis set will include all randomized participants who receive at least one dose (partial or full) of the study intervention, and participants will be analyzed based on the treatment they receive, regardless of the treatment group to which they are assigned.

[0220] The pharmacokinetic analysis set included all participants who received at least one full dose of guselkumab and had at least one observed post-dose PK data.

[0221] The immunogenicity analysis set included all participants who received at least one dose of guselkumab and had at least one observed post-dose immune response data.

[0222] Efficacy analysis Most data will be summarized using simple descriptive summary statistics such as n, mean, SD, median, interquartile range, minimum, and maximum for continuous variables, and counts and percentages for discrete variables.

[0223] For the primary endpoint (change from baseline in mRSS at week 24), treatment comparisons will be performed using a mixed-effects model repeated measures (MMRM) model. The MMRM model includes treatment group, baseline mRSS, stratification factors, visit, treatment group by visit interaction, and baseline mRSS by visit interaction as fixed effects. Treatment effects will be estimated based on least squares (LS) means of differences. p-values ​​for LS mean differences with two-sided 80% CIs will be presented. Sensitivity and subgroup analyses for the primary endpoint will be performed, where appropriate. Details of these analyses and data processing rules will be specified in the statistical analysis plan (SAP).

[0224] All other efficacy endpoints will be summarized over time by treatment group. Treatment comparisons will be performed using MMRM models where there are repeated continuous measures, or logistic models where there are dichotomous response variables. No adjustments for multiple comparisons for secondary endpoints will be made, and all p-values ​​will be considered nominal. Detailed methods of analysis and data processing rules will be provided in SAP.

[0225] Safety analysis Routine safety evaluations will be based on the safety analysis set. Adverse events, SAEs, related AEs, and AEs by severity will be summarized by treatment group. Further specification of other AESIs such as ILD will be described in the SAP.

[0226] Laboratory parameters and changes from baseline in selected laboratory parameters (hematology and chemistry), as well as the number of participants with abnormal laboratory parameters (hematology and chemistry) based on National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE) toxicity grading, will be summarized by treatment group.

[0227] Clinically relevant ECG abnormalities will be assessed by frequency tables. Descriptive statistics of temperature, pulse / heart rate, respiratory rate, and blood pressure (systolic and diastolic) values ​​and changes from baseline will be summarized at each scheduled time point. The proportion of participants with values ​​above clinically significant limits will be summarized.

[0228] Pharmacokinetic analysis Unless otherwise stated, PK analyses are based on the PK analysis set. Serum guselkumab concentrations over time are summarized using descriptive statistics at each nominal sampling time point. All concentrations below the lowest quantifiable concentration or missing data will be labeled as such in the concentration database or data presentation. Concentrations below the lowest quantifiable concentration will be treated as zero in summary statistics.

[0229] Detailed rules for the analysis, including exclusions from the PK analysis, will be specified in the SAP.

[0230] Population PK modeling may be performed, where appropriate. If population PK analyses are performed, the results of these analyses will be presented in a separate report.

[0231] Immunogenicity analysis The incidence and titers of anti-guselkumab antibodies will be summarized for the immunogenicity analysis set. A list of participants testing positive for anti-guselkumab antibodies will be provided. Maximum titers of anti-guselkumab antibodies will be summarized for participants testing positive for antibodies to guselkumab.

[0232] The incidence of Nabs to guselkumab will be summarized for participants who tested positive for antibodies to guselkumab and had a sample evaluable for neutralizing antibodies to guselkumab.

[0233] Pharmacodynamic analysis Serum will be collected from all subjects to assess PD markers associated with guselkumab and markers associated with SSc. Measurements may include, but are not limited to, serum levels of IL-17A, IL-17F, IL-22, beta defensin-2 (BD-2), and SAA. More extensive proteomic profiling (e.g., by Olink technology) for biomarker discovery can be performed.

[0234] Blood samples for isolation of PBMCs are also collected for subsequent immunophenotyping analysis by multiparameter flow cytometry or mass cytometry (CyTOF) analysis to measure immune cell populations before and during treatment. Gene expression analysis can also be performed, which may include single-cell RNA sequencing (RNA-seq) profiling.

[0235] The results of the pharmacodynamic / biomarker analysis may be presented in a separate report.

[0236] Biomarker analysis Characterization of gene expression changes in the skin during treatment will be determined and analyzed by RNA-seq at weeks 0 and 24. If feasible, non-lesional (uninfected) skin samples will be taken at week 0 and lesional (week 0) samples will be taken at both weeks 0 and 24. If feasible, characterization of histoimmunopathological changes in the skin as determined by immunohistochemistry (IHC) / immunofluorescence (IF) / in situ hybridization (ISH) as well as histological analysis (e.g., fibrosis) may be performed. Multiparameter protein profiling (e.g., recall CyTOF) may be considered.

[0237] Pharmacokinetic / pharmacodynamic analysis Relationships between serum concentrations of guselkumab and efficacy measures, including biomarkers in blood or skin biopsies, and / or relevant PD endpoints can be explored graphically where appropriate. After any visual trends are observed, additional analyses may be performed if deemed necessary.

[0238] Pharmacogenomics Analysis Genetic analysis may help to identify population subgroups that respond differently to a drug. A single sample at week 0 will be used to explore genetic factors that may affect the molecular effect, clinical efficacy, or tolerability of guselkumab and to identify genetic factors associated with SSc. The locus of interest is the IL12RB1 gene, where reported SSc risk alleles have been identified and there is a potential association with the expression of the gene product (encoding the receptor for IL-23). ​​Analysis applied to the DNA sample may focus on genotyping using molecular arrays to scan for polymorphisms across the genome. Participation in DNA sampling is voluntary.

[0239] Study population Screening of eligible participants will occur within 6 weeks prior to administration of the study intervention. The inclusion and exclusion criteria for enrolling participants in this study are described below. If there are questions about these criteria, investigators must consult with the appropriate sponsor personnel and resolve the issue prior to enrolling the participant in the study. Deviations will not be permitted.

[0240] 1.1. Inclusion criteria Each potential participant must meet all of the following criteria to be enrolled in the study: 1. Male or female (according to the reproductive organs and functions assigned by the chromosomal complement). 2. Age 18-75 years (inclusive).

[0241] Participant types and disease characteristics 3. Medically stable based on physical examination, medical history, vital signs, and a 12-lead ECG performed at screening. Any abnormalities must be consistent with underlying disease in the study population, and this determination must be documented in the participant's source documentation and initialized by the investigator. 4. Medically stable based on clinical laboratory tests performed at screening. If the results of a serum chemistry panel, including liver enzymes, coagulation, hematology, or urinalysis, are outside the normal reference range, the participant may be included only if the investigator determines the abnormal results or deviations from normal results to be not clinically significant or appropriate and reasonable for the population under study. This determination must be recorded on the participant's source document and initialed by the Investigator. 5. Diagnosis of SSc according to ACR and EULAR 2013 criteria. 6. Diffuse cutaneous SSc according to LeRoy criteria, i.e., dermal fibrosis proximal to the elbows and knees in addition to acral fibrosis. 7. Disease duration (defined as the time from the first non-Raynaud's phenomenon episode) ≤36 months. 8. ≥10 and ≤22 mRSS units at Screening and Week 0. 9. FVC ≥ 60% of predicted at screening. 10. DLCO (corrected for hemoglobin) ≥ 40% of predicted at screening. 11. Participants who meet one of the following criteria at screening: a) An increase of 3 mRSS units or more compared to an evaluation performed within the past 2-6 months. * b) Complications in one new body area with an increase of ≥2 mRSS units compared to evaluations performed within the past 2–6 months. * c) complications in two new body regions with an increase of 1 mRSS unit or more compared to evaluations within the previous 2-6 months. * * Exclude participants who experienced a decrease in mRSS within 2 months. 12. Standards revised by Amendment 1 12.1 With laboratory screening test results within the following parameter ranges, one retest of the laboratory values ​​is permitted if one or more of the laboratory parameters are out of range:

[0242] [Table 4]

[0243] Concomitant or past drug therapy received 13. Regular treatment with the following medications without changes in dose or frequency is permitted as follows: a) Oral glucocorticoids (mean daily dose of prednisolone or equivalent ≤10 mg) At least 6 weeks prior to the first dose of study intervention and at least 2 weeks on a stable dose. If you are not currently using oral glucocorticoids They must not have received any of the study interventions for more than 6 weeks prior to the first dose. b) Stable doses of nonsteroidal anti-inflammatory drugs (NSAIDs) or other analgesics ≥2 weeks prior to the first dose of study intervention. c) A stable dose of an endothelin receptor antagonist (e.g., bosentan, ambrisentan, and macitentan) for ≥2 weeks prior to the first dose of the study intervention. d) Phosphodiesterase 5 inhibitors (e.g., sildenafil and tadalafil) at a stable dose for ≥2 weeks prior to the first dose of the study intervention. e) Prostacyclin (e.g., epoprostenol, iloprost, and treprostinil) at a stable dose ≥2 weeks prior to the first dose of study intervention. f) Permitted topical medication for skin conditions ≥4 weeks prior to the first dose of study intervention.

[0244] tuberculosis 14. Subjects will be considered eligible according to the following TB screening criteria: a) No history of latent or active TB prior to screening. Exceptions are made for participants with a history of latent TB who meet one of the following criteria: - currently receiving treatment for latent TB, or - Initiate treatment for latent TB before first dose of study intervention or - Documented completion of adequate treatment for latent TB within 5 years prior to first administration of the intervention. It is the investigator's responsibility to verify the adequacy of prior anti-TB treatment and provide appropriate documentation. Patients with history and documentation of completion of adequate treatment for latent TB more than 5 years prior to first administration of the study intervention are not eligible. b) No signs or symptoms suggestive of active TB upon medical history and / or physical examination. c) Have not had recent close contact with anyone with active TB or, if such contact has occurred, be referred to a TB specialist for further evaluation and, if warranted, receive appropriate treatment for latent TB prior to the first dose of the study intervention. d) Have a negative interferon-gamma release assay (IGRA; QuantiFERON-TB® or T-SPOT®) result within 2 months prior to the first dose of study intervention, or have a newly identified positive IGRA test result indicating that active TB has been ruled out and appropriate treatment for latent TB has been initiated prior to the first dose of study intervention (see Clinical Laboratory Manual). Subjects with inconclusive results from the first IGRA test should have the study repeated. If the results of the second IGRA test are also inconclusive, the subject may be enrolled without treatment for latent TB if active TB has been ruled out, the chest radiograph shows no abnormalities suggestive of TB (active or old, inactive TB), and the subject has no further risk factors for TB as determined by the investigator. This determination should be promptly reported to the sponsor's medical monitor, recorded in the subject's source documentation, and abbreviated and initialed by the investigator. NOTE: Participants with a history of latent TB and who are currently on treatment for latent TB or have documentation of having completed adequate treatment as described above do not require an IGRA at screening, and participants who demonstrate completion of adequate treatment as described above are not required to start further treatment for latent TB. e) Chest radiographs (both anteroposterior and lateral) taken within 12 weeks prior to the first dose of study intervention, read by a qualified physician (e.g., radiologist or pulmonologist) and showing no evidence of currently active or older inactive TB. Chest computed tomography (CT) may also be performed if deemed appropriate by the investigator.

[0245] Gender and contraception / barrier requirements 15. Females of childbearing potential must have a negative urine pregnancy test at screening and baseline. 16.Women must fulfil the following conditions: A. There is no possibility of pregnancy b. There is a possibility of pregnancy, and Implement a highly effective, preferably user-independent, method of contraception (with a failure rate of <1% per year if used consistently and accurately) and agree to maintain a highly effective method while receiving the intervention and until 12 weeks after the last dose (i.e., end of relevant systemic exposure). The investigator should assess the possibility of contraceptive failure (e.g., noncompliance, recent initiation) in relation to the first dose of the study intervention. Note: If a participant's fertility changes after study initiation (e.g., a premenarcheal woman experiences menarche) or pregnancy risk changes (e.g., a sexually inactive woman becomes sexually active), women must begin using highly effective contraception as described throughout the inclusion and exclusion criteria. 17. Women must agree not to donate eggs (egg cells, oocytes) for assisted reproductive purposes during the study and for 12 weeks after the last dose of the study intervention. 18. Standards revised by Amendment 1 18.1 Male participants must agree to the following during the study and for at least 12 weeks after the last dose of study intervention: ● You must agree not to donate sperm for reproductive purposes. ● Male participants must wear a condom when engaging in any act of transferring ejaculate to another person. • Male participants should also be counseled about the benefits of their female partners using a highly effective method of contraception, as condoms can break or leak.

[0246] Informed consent 19. Each person must sign an informed consent form (ICF) indicating that they understand the purpose of the study and the procedures it will entail, and are willing to participate in the study. 20. If you agree to provide a voluntary DNA sample for research, you must sign a separate informed consent form. Refusal to consent for a voluntary DNA research sample will not exclude you from participating in the study. 21. Be willing and able to adhere to the lifestyle restrictions specified in this protocol.

[0247] 1.2. Exclusion criteria Potential participants who meet any of the following criteria will be excluded from participating in the study:

[0248] Medical conditions 1. History of hepatic or renal failure (estimated creatinine clearance <60 mL / min); or significant cardiac, vascular, pulmonary, gastrointestinal, endocrine, neurological, hematological, rheumatologic, psychiatric, or metabolic disorder. 2. Any known severe or uncontrolled SSc complication, including hemoptysis, pulmonary hemorrhage, or renal crisis. Have severe pulmonary hypertension as determined by echocardiography and pulmonary function testing or right heart catheterization. Severe pulmonary hypertension includes, but is not limited to: Left ventricular ejection fraction < 40% Peak tricuspid regurgitation velocity >3.4 (m / s) ● Resting heart rate is less than 50 bpm ● Systolic blood pressure is less than λ0mmHg ECG findings suggesting significant conduction abnormalities 3. Patients with interstitial lung disease requiring oxygen therapy. 4. Having any rheumatic disease other than SSc that may interfere with the evaluation of SSc, such as RA, polymyalgia rheumatica (PMR), systemic lupus erythematosus, polymyositis / dermatomyositis, etc. 5. Currently has a diagnosis or signs or symptoms of a serious, advanced, or uncontrolled renal, cardiac, vascular, pulmonary, gastrointestinal, endocrine, neurological, hematological, rheumatic, psychiatric, or metabolic disorder (or any other concomitant medical condition that, in the opinion of the Investigator, would put the participant at risk by participating in this study). 6.Currently have or have had any major ischemic event within 12 weeks of first study intervention. 7. Have a history of lymphoproliferative disorders, including lymphoma; have MGUS, monoclonal gammopathy of undetermined significance; or have signs and symptoms suggestive of a possible lymphoproliferative disorder, such as lymphadenopathy or splenomegaly. 9. History of or ongoing chronic or recurrent infections, including but not limited to chronic kidney infections, chronic chest infections (e.g., bronchiectasis), recurrent urinary tract infections (recurrent pyelonephritis or chronic persistent cystitis), fungal infections (e.g., mucocutaneous candidiasis, but excluding fungal infections of the nail bed), or open, draining, or infected skin wounds or ulcers. 10. Subject currently has or has had a serious infection (including sepsis, pneumonia, or pyelonephritis) or has been hospitalized or received IV antibiotics for an infection within 2 months prior to the first study intervention. 11. Current or past nontuberculous mycobacterial infection or clinically significant opportunistic infection (e.g., cytomegalovirus, pneumocystis, invasive aspergillosis). 12. History of latent or acute granulomatous infection (including histoplasmosis or coctidioidomycosis) prior to screening. 13. Have a history of an infected joint prosthesis or have received antibiotics for a suspected infection of the joint prosthesis within 6 months of the first study intervention if the prosthesis has not been removed or replaced. 14. Have a history of or are infected with the human immunodeficiency virus (HIV [serologically positive for HIV antibodies]); have tested positive for Hepatitis B virus (HBV) infection; or have antibodies to Hepatitis C virus (HCV) at screening. 15. During the 6 weeks prior to baseline, had either: (a) confirmed SARS-CoV-2 (coronavirus disease 2019 [COVID-19]) infection (positive test), or (b) suspected SARS-CoV-2 infection (clinical features without documented test results), or (c) close contact with a person with known or suspected SARS-CoV-2 infection. Exceptions to this criterion may be given if the participant has a documented negative result for a validated SARS-CoV-2 test: (i) acquired at least 2 weeks after conditions (a), (b), or (c) above (timed from the resolution of significant clinical features, if present, e.g., fever, cough, and dyspnea); and (ii) All of the above conditions (a), (b), and (c) are absent during the period between the negative test result and the baseline testing visit. Notes on COVID-related exclusions: ● The field of COVID-related testing (for the presence of and immunity to the SARS-CoV-2 virus) is rapidly evolving. Additional testing, if deemed necessary by the Investigator, may be performed as part of screening and / or during the study in accordance with current regulations / guidance / standard of care from authorities. ● Note: For those who may be at higher risk for severe COVID-19 illness, follow guidance from local health authorities when weighing the potential benefits and risks of enrolling in the trial and during your participation in the trial. 16. Have experienced a recent monodermatomal zoster rash within the past 4 months. Have ever had multidermal zoster (defined as lesions appearing outside of the primary or adjacent dermatomes) or a central nervous system zoster infection. 17.Currently have a malignancy or have a history of malignancy within 5 years prior to screening (excluding non-melanoma skin cancer that has been adequately treated and has had no evidence of recurrence for at least 3 months prior to the first study intervention, or cervical intraepithelial neoplasia that has been treated and has had no evidence of recurrence for at least 3 months prior to the first study intervention). NOTE: Pre-malignant lesions should be discussed with the sponsor's medical monitor. 18.Has had a transplanted organ, including HSCT (excluding corneal transplant surgery >3 months prior to the first study intervention). 19. Known allergy, hypersensitivity, or intolerance to guselkumab or its excipients.

[0249] Diagnostic evaluation 20. Has had major surgery (e.g., requiring general anesthesia and hospitalization) within 8 weeks prior to screening or has not fully recovered from such surgery, or has such surgery planned during the period in which the participant is expected to participate in the study. NOTE: Participants who are scheduled for surgical treatment performed under local anesthesia are eligible to participate. 21. History of drug or alcohol abuse within 1 year prior to screening according to the criteria in the Diagnostic and Statistical Manual of Disorders (5th ed.).

[0250] Previous and Concomitant Therapies 22.Has had prior treatment with any investigational or approved immunomodulatory biologic agent prior to the first study intervention, including but not limited to: ● Received the following treatment: - IL-23 inhibitor therapy (including but not limited to guselkumab, risankizumab, tildrakizumab, brazikumab, mirikizumab) - IL-12 / 23 inhibitor (ustekinumab) - IL-17 inhibitors (secukinumab, ixekizumab, brodalumab) - Have received any of the following treatments within 12 weeks or 5 half-lives (whichever is longer): - Tocilizumab (Actemra) - Sirikumab, Sarilumab, Mavrilimumab, Abatacept, Belimumab - Tyrosine kinase inhibitors (nintedanib) - Systemic and topical Janus kinase inhibitors (e.g., tofacitinib, upadacitinib) - Received the following treatments within 1 year or 5 half-lives (whichever is longer): - Rituximab Note: Any biologic agents not listed should be discussed and agreed with the sponsor's medical monitor. 23. Received the following treatments: Within 6 months of study intervention: - Cytotoxic drugs (cyclophosphamide, chlorambucil, nitrogen mustard, or other alkylating agents) - Intravenous immunoglobulin (IVIG), apheresis therapy (plasmapheresis or leukapheresis) Within 6 weeks of the first study intervention: - Systemic immunosuppressants (including but not limited to cyclosporine A, azathioprine, tacrolimus, sirolimus, sulfasalazine, leflunomide or mycophenolate mofetil / mycophenolate with cholestyramine washout) - Intramuscular, intra-articular, intrasynovial, epidural, intralesional, or IV glucocorticoids Any questions or concerns regarding the use of these therapies should be discussed with the sponsor and / or medical monitor.

[0251] Previous / concurrent clinical trial experience 24.Has received an investigational intervention (including an investigational vaccine) within 3 months or 5 half-lives (whichever is longer), or has used an invasive investigational medical device within 3 months of the first planned dose of the investigational intervention, or is currently enrolled in a clinical trial. 25. Received a Bacille Calmette-Guerin (BCG) vaccination within 12 months of randomization or any other live bacterial or live viral vaccination within 12 weeks of randomization. 26.Women who are pregnant, breastfeeding, or planning to become pregnant while enrolled in this study or within 12 weeks after administration of the last study intervention. 27.Men who plan to father a child while enrolled in this study or within 12 weeks after administration of the last study intervention. 28. Any condition where, in the opinion of the investigator, participation would not be in the participant's best interests (e.g., would compromise their health) or which might prevent, limit, or confound the evaluations specified in the protocol.

[0252] Study intervention The sponsor-manufactured combination product for use in this study is a pre-filled syringe (PFS) assembled with an UltraSafe Plus™ passive needle guard (PFS-U).

[0253] Preparation / Handling / Storage Guselkumab final vialed product (FVP [IV]) for IV administration will be supplied as a sterile solution in single-use glass vials containing 20 mL at a concentration of 10 mg / mL. Study interventions will be prepared for IV administration based on instructions provided to IPPI clinical sites.

[0254] Guselkumab will be supplied as a 100 mg / mL sterile solution in a single-use PFS (PFS-U) assembled in an UltraSafe Plus™ passive needle guard for SC administration. Guselkumab placebo will be supplied as a 1 mL sterile solution in a single-use PFS assembled in a PFS-U for SC administration.

[0255] Guselkumab and guselkumab placebo should be a clear, colorless to pale yellow solution that may contain small translucent particles. Do not use guselkumab or guselkumab placebo if the liquid is cloudy, discolored, or has large particles. Light protection is not required during preparation and administration of the study intervention, but direct exposure to sunlight must be avoided. Aseptic technique should be used during preparation and administration of study intervention materials.

[0256] Example 2 - Results of treatment of patients with SSc Three cases of psoriasis vulgaris (pSV) complicated with SSc were treated with guselkumab at the Systemic Sclerosis Center of the University of Tokyo Hospital in Japan, and good therapeutic effects were obtained not only against pSV but also against SSc. All cases were suspected to have pSV by local physicians, who referred them for detailed examination and treatment. A detailed examination including pathology confirmed the diagnosis of pSV in these patients. Furthermore, these patients had skin sclerosis extending from the fingers to the upper arms. All cases were judged to have SSc complicated based on the ACR / EULAR criteria. The time of the first appearance of SSc symptoms other than Raynaud's phenomenon was identified by questioning, and the duration of SSc disease was determined. On further examination, all patients were found to have gastroesophageal reflux disease (ERD) by gastrointestinal endoscopy. Furthermore, one patient (Case #2; Table 4) had mild interstitial lung disease (ILD) on HRCT with percent predicted forced vital capacity (FVC) and diffusing capacity for carbon monoxide (DLco) of 74.7% and 88.1%, respectively. The absence of other complications, including arthritis, pulmonary hypertension, and renal crisis, was confirmed by a rheumatologist. After these systemic evaluations, all patients wished to be treated with a biologic for pSV. For this purpose, 100 mg of GUS was administered subcutaneously at weeks 0 and 4, and then at 8-week intervals. In all cases, the only treatment other than GUS was topical steroids. pSV severity was assessed by the psoriasis area and severity index (PASI), and SSc was assessed by the modified Rodnan total skin thickness score (MRTSS) and the combined response index for SSc (CRISIS). Patient demographic information and clinical data at the start of and 6 months after treatment with GUS are shown in Table 4.

[0257] In all cases, treatment with GUS resulted in zero PASI after 6 months of treatment. SSc skin sclerosis was also improved, and the MRTSS was reduced by more than 6 in each case. CRISS was also improved in all cases. Furthermore, GERD was improved after GUS administration, and the F scale, a measure of severity, was reduced by more than 5 points. There was no worsening of ILD throughout the treatment period. No adverse events were observed. Written informed consent was obtained from all patients. This report suggests that anti-IL-23 antibody therapy for SSc may be beneficial. The effects of anti-IL-23 antibody therapy for SSc are expected to become clear in future large-scale trials.

[0258] [Table 5] dcSSc, diffuse cutaneous SSc; lcSSc, limited cutaneous SSc; WBC, white blood cells; Hb, hemoglobin; PLT, platelets; CRP, C-reactive protein; SP-D, surfactant protein D; KL-6, Krevs von den Lungen-6; RNAP, anti-RNA polymerase III antibody; Topo I, anti-topoisomerase I antibody; CENP, anti-centromere protein antibody; +, present; -, absent.

[0259] The present invention can be described with reference to the following numbered embodiments. 1. Use of an antibody specific for IL23 for the treatment of systemic sclerosis (SSc) in a patient, the antibody comprising a light chain variable region and a heavy chain variable region, 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 comprising the CDRL3 amino acid sequence of SEQ ID NO:6, The heavy chain variable region is 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 3, and wherein the use results in a clinical response in a patient. 2. The use of embodiment 1, wherein the antibody is administered at a first dose, at a dose about 4 weeks after the first dose, and at a dose about 8 weeks after the first dose. 3. The use of embodiment 2, wherein the antibody is administered intravenously. 4. The use of embodiment 1, wherein the first dose and the dose about 4 weeks after the first dose and the dose about 8 weeks after the first dose are 200 mg or 400 mg of antibody. 5. The use of embodiment 4, wherein the initial dose and the dose about 4 weeks and about 8 weeks after the initial dose are 400 mg of antibody. 6. The use of embodiment 5, wherein the antibody is administered subcutaneously about every 4 weeks, following administration about 8 weeks after the first dose. 7. The use of embodiment 6, wherein the subcutaneous administration is 200 mg of antibody. 8. Patients are identified as responders to the antibody and meet clinical endpoints, which are change from baseline in mRSS, worsening of mRSS, American College of Rheumatology Composite Response Index (ACR) in dcSSc. 2. The use of embodiment 1, wherein the objective of the present invention is to achieve a score of 0.6 on the Critical Isolation Stress Test (CRISS), change from baseline in forced vital capacity (FVC) and percent predicted FVC, change from baseline in measured absolute diffusing capacity of the lung for carbon monoxide (DLCO) and calculated percent predicted DLCO, change from baseline in number of digital ulcers in patients with digital ulcers at baseline, change from baseline in Health Assessment Questionnaire-Disability Index (HAQ-DI) score, change from baseline in Frequency Scale of Gastroesophageal Reflux Symptoms (FSSG) score, change from baseline in fibrotic changes and worsening of fibrotic changes as assessed by high-resolution computed tomography (HRCT), and change from baseline in capillaroscopic assessment in the Patient Global Assessment (PGA) and / or Physician Global Assessment (PhGA). 9. The use of embodiment 8, wherein the clinical endpoint is measured at about 24 weeks, 52 weeks, and / or 104 weeks after the initial treatment. 10. The use of embodiment 9, wherein the clinical endpoint is measured about 24 weeks after the initial treatment. 11. The use of embodiment 1, wherein the antibody comprises the light chain variable region amino acid sequence of SEQ ID NO:8 and the heavy chain variable region amino acid sequence of SEQ ID NO:7. 12. The use according to embodiment 1, wherein the antibody comprises a light chain amino acid sequence of SEQ ID NO: 10 and a heavy chain amino acid sequence of SEQ ID NO: 9. 13. The use according to embodiment 11 or 12, wherein the antibody is present in a composition comprising 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, and 0.053% (w / v) polysorbate 80 of the pharmaceutical composition, and the diluent is water at standard conditions. 14. The use of embodiment 1, further comprising the use of one or more additional drugs used to treat SSc. 15. The use of embodiment 14, wherein the additional drug is selected from the group consisting of immunosuppressants, nonsteroidal anti-inflammatory drugs (NSAIDs), methotrexate (MTX), anti-B cell surface marker antibodies, anti-CD20 antibodies, rituximab, TNF inhibitors, corticosteroids, and costimulatory regulators. 16. Use of an antibody specific for IL23 for the treatment of moderately to severely active SSc in a patient by administering: (i) an initial intravenous dose of 400 mg; (ii) an intravenous dose of 400 mg about 4 weeks after the initial dose; (iii) an intravenous dose of 400 mg about 8 weeks after the initial dose; and (iv) an intravenous dose about 8 weeks after the initial dose, followed by subcutaneous administration of 200 mg of the antibody about every 4 weeks, The antibody comprises a light chain variable region amino acid sequence of SEQ ID NO:8 and a heavy chain variable region amino acid sequence of SEQ ID NO:7, and the patient is a responder to the antibody by being identified as meeting a clinical endpoint approximately 24 weeks after the first administration, and the clinical endpoint is change from baseline in modified Rodnan Skin Score (mRSS).

[0260] Sequence Listing

[0261] TIFF2025500882000007.tif209132

[0262] TIFF2025500882000008.tif214132

[0263] TIFF2025500882000009.tif196160

[0264] TIFF2025500882000010.tif221132

[0265] TIFF2025500882000011.tif217132

[0266] TIFF2025500882000012.tif162132

Claims

1. 1. A method of treating systemic sclerosis (SSc) in a patient, comprising administering to the patient an antibody specific for IL23; The antibody comprises a light chain variable region and a heavy chain variable region, 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 comprising the CDRL3 amino acid sequence of SEQ ID NO:6, The heavy chain variable region 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 comprising the CDRH3 amino acid sequence of SEQ ID NO:3, The method, wherein said patient is considered a responder to said antibody, said patient being a responder to said antibody.

2. 10. The method of claim 1, wherein the antibody is administered at a first dose, at about 4 weeks after the first dose, and at about 8 weeks after the first dose.

3. 3. The method of claim 2, wherein the first administration, the administration about 4 weeks after the first administration, and the administration about 8 weeks after the first administration is about 200 mg or about 400 mg of the antibody.

4. 4. The method of claim 3, wherein the antibody is administered intravenously for the first administration, the administration about 4 weeks after the first administration, and the administration about 8 weeks after the first administration.

5. 5. The method of claim 4, further comprising administering said administration about 8 weeks after said initial administration, followed by administration of a maintenance administration of said antibody about every 4 weeks.

6. 6. The method of claim 5, wherein the maintenance dose is about 200 mg of antibody and is administered subcutaneously.

7. 2. The method of claim 1, wherein the patient is a responder to the antibody by being identified as meeting a clinical endpoint.

8. 8. The method of claim 7, wherein the clinical endpoint is change from baseline in modified Rodnan Skin Score (mRSS).

9. 9. The method of claim 8, wherein the clinical endpoint is measured about 24 weeks after the first administration.

10. 8. The method of claim 7, wherein the clinical endpoint is selected from the group consisting of: change from baseline in mRSS, worsening of mRSS, achieving a score of 0.6 on the American College of Rheumatology Composite Response Index (ACR CRISS) in dcSSc, change from baseline in forced vital capacity (FVC) and percent predicted FVC, change from baseline in measured absolute diffusing capacity of the lung for carbon monoxide (DLCO) and calculated percent predicted DLCO, change from baseline in number of digital ulcers in patients with digital ulcers at baseline, and change from baseline in Health Assessment Questionnaire-Disability Index (HAQ-DI) score.

11. 11. The method of claim 10, wherein the clinical endpoint is measured at about 24 weeks, 52 weeks, and / or 104 weeks after initial treatment.

12. 12. The method of claim 11, wherein the clinical endpoint is measured about 24 weeks after initial treatment.

13. The method of claim 1 , wherein the antibody comprises a light chain variable region amino acid sequence of SEQ ID NO:8 and a heavy chain variable region amino acid sequence of SEQ ID NO:

7.

14. The method of claim 1 , wherein the antibody comprises a light chain amino acid sequence of SEQ ID NO:10 and a heavy chain amino acid sequence of SEQ ID NO:

9.

15. 15. The method of claim 13 or 14, wherein the antibody is present in a composition comprising 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, and 0.053% (w / v) polysorbate 80 of the pharmaceutical composition, and the diluent is water at standard conditions.

16. 10. The method of claim 1, further comprising administering to the patient one or more additional drugs used to treat SSc.

17. 17. The method of claim 16, wherein the additional drug is selected from the group consisting of immunosuppressants, nonsteroidal anti-inflammatory drugs (NSAIDs), methotrexate (MTX), anti-B cell surface marker antibodies, anti-CD20 antibodies, rituximab, TNF inhibitors, corticosteroids, and costimulatory regulators.

18. 1. A method of treating SSc in a patient, comprising: (i) administering to a patient an initial intravenous administration of 400 mg of an IL23 specific antibody; (ii) administering intravenously 400 mg of the antibody about 4 weeks after the initial administration; (iii) administering intravenously 400 mg of the antibody about 8 weeks after the initial administration; and iv) administering subcutaneously 200 mg of the antibody about every 4 weeks thereafter, the antibody comprises a light chain variable region amino acid sequence of SEQ ID NO:8 and a heavy chain variable region amino acid sequence of SEQ ID NO:7, and the patient is a responder to the antibody by being identified as meeting a clinical endpoint about 24 weeks after the first administration; A method wherein the clinical endpoint is change from baseline in modified Rodnan Skin Score (mRSS).

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