Safe and effective method for treating psoriasis by anti il-23 specific antibody

Abstract

To provide a method for treating psoriasis of a patient.SOLUTION: An IL-23 specific antibody that is Guselkumab for example is administered at an effective amount having clinically proven safety. A patient achieves scores of PASI 90, PASI 100, or IGA 0 or 1 which are measured on 16-th week, 24-th week, 32-nd week, 40-th week, and 48-th week after a first treatment, and realizes higher effectiveness than a patient treated by Secukinumab antibody.SELECTED DRAWING: None

Description

[Technical field]

[0001] (Reference to electronically submitted sequence listing) This application has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. A copy of this ASCII file is available at It was created on the 19th of the month and the file name is PCTSEQUENCELISTING.txt. Its size is 80,004 bytes.

[0002] FIELD OF THEINVENTION The present invention relates to a method for treating psoriasis with an antibody that binds to human IL-23 protein. Specifically, the present invention relates to an anti-IL-23 specific antibody and a method for treating psoriasis in patients. Administering certain pharmaceutical compositions of antibodies (e.g., guselkumab) is safe and effective for This relates to a method for [Background technology]

[0003] Interleukin (IL)-12 is a glycosyltransferase that binds two disulfide bonds. The oligomerized protein subunits (denoted p35 and p40 because of their approximate molecular weight) IL-12 is a secreted heterodimeric cytokine composed of the IL-12 complex (IL-13) and IL-14 (IL-15). It is mainly produced by antigen-presenting cells, and is activated by T cells or natural killer (N K) Stimulates cell-mediated immunity by binding to a two-chain receptor complex expressed on the surface of cells. The IL-12 receptor β-1 (IL-12Rβ1) chain promotes the expression of the p40 subunit of IL-12. However, IL-12 binds to the mitochondrial nucleus, providing the primary interaction between IL-12 and its receptor. These results suggest that the IL-1 receptor signaling pathway is essential for the activation of receptor-bearing cells. It confers IL-12p35 ligation of the second receptor chain, IL-12Rβ2. (Presky et al., 1996). IL-12 signaling in parallel with antigen presentation are associated with a T helper 1 (Th1) phenotype characterized by interferon-γ (IFNγ) production. It is thought that this induces T cell differentiation toward the immune system (Trinchieri, 2003). Th1 cells promote immunity to some intracellular pathogens and produce complementary antibody isotypes. IL-12 is thought to contribute to tumor immune surveillance by promoting the production of IL-12 in host defense immune cells. It is believed to be an important component of the immune system.

[0004] The p40 protein subunit of IL-12 also binds to a separate protein designated p19. It has been discovered that IL-23 can associate with protein subunits to form a novel cytokine, IL-23. IL-23 also signals through a two-chain receptor complex. The p40 subunit is shared between IL-12 and IL-23, The IL-12Rβ1 chain is also shared between IL-12 and IL-23. L-23-specific intracellular signaling (e.g., STAT3 phosphorylation) and subsequent T cell The second component of the IL-23 receptor complex, IL-23R, confers IL-17 production by the IL-23 receptor. The IL-23p19 nucleotide sequence is a structural component of the IL-23p19 nucleotide sequence (Parham et al., 2002; Aggar Recent studies have demonstrated that IL-23 plays a key role in the regulation of IL-12 production and function. Despite the structural similarities between these two cytokines, the biological functions are distinct. showed that (Langrish et al., 2005).

[0005] Neutralization of IL-12 by antibodies has been shown to be effective in treating psoriasis, multiple sclerosis (MS), and Rheumatoid arthritis, inflammatory bowel disease, insulin-dependent (type 1) diabetes mellitus, and uveitis Abnormal regulation of IL-12 and Th1 cell populations for efficacy in treating animal models It has been associated with many immune-mediated diseases (Leonard et al., 1995; H ong et al. (1999), Malfait et al. (1998), Davidson et al. (19 However, these studies targeted the shared p40 subunit, Both IL-12 and IL-23 were neutralized in vivo. Whether L-23 mediated disease or whether both sites were involved in the pathogenesis of disease suppression was unclear. It is unclear whether inhibition of IL-23 is necessary to achieve the anti-inflammatory effect. The strategy may provide benefits equivalent to those of the IL-12p40 strategy in IL-23p19-deficient or was confirmed by specific antibody neutralization of IL-23 (Cua et al., 2003; Murphy et al., 2004). (2003), Benson et al. (2004). There is growing evidence regarding the specific role of IL-23 in inhibiting the IL-12 pathway. Neutralizing IL-23 without any treatment has limited impact on important host defense immune mechanisms. This could provide an effective therapy for immune-mediated diseases, which is in contrast to current treatment options. would represent a significant improvement.

[0006] Psoriasis is associated with a number of health problems, including psoriatic arthritis (PsA), depression, cardiovascular disease, and hypertension. Common chronic conditions with significant comorbidities such as hypertension, obesity, diabetes, metabolic syndrome, and Crohn's disease It is an immune-mediated skin disease. Plaque psoriasis is the most common form of the disease and is characterized by silvery white spots. The lesions are pruritic, painful, and disfiguring. Psoriasis is often disabling and disabling, with a significant proportion of people with psoriasis experiencing damage to the hands / nails, face, feet, and genitals. Therefore, psoriasis significantly affects health-related quality of life (HRQoL). These adverse effects can extend beyond physical skin symptoms to include impaired daily activities. For example, psoriasis imposes physical and psychosocial burdens on the body that can interfere with the treatment of certain conditions. High levels of negative impact on family, spouse, social and professional relationships It has been linked to increased incidence of depression and suicidal tendencies.

[0007] Histological characterization of psoriatic lesions reveals psoriasis caused by abnormal keratinocyte proliferation and differentiation. Skin thickening and dermal infiltration and colocalization of CD3+ T lymphocytes and dendritic cells were evident. The pathogenesis of psoriasis has not been fully elucidated, but genetic and protein analysis have been reported. Analysis revealed that IL-12, IL-23, and their downstream molecules are overexpressed in psoriatic lesions. It has been shown that some of these may correlate with the severity of psoriasis disease. Some of the therapies being studied have been shown to have IL-12 and IL-2 inhibitors, which may contribute to their efficacy. 3. Th1 and Th17 cells regulate the levels of vasodilators, chemoattractants, and They produce effector cytokines that induce the production of substances and the expression of adhesion molecules on endothelial cells. These in turn can result in monocyte and neutrophil recruitment, T cell infiltration, angiogenesis, and endothelial cell proliferation. Activated keratinocytes promote the activation and hyperplasia of neutrophils, They can produce chemoattractant factors that promote the trafficking of monocytes, T cells, and dendritic cells. Thus, a cycle of inflammation and keratinocyte hyperproliferation is established.

[0008] The etiology of psoriasis is based on tumor necrosis factor-α (TNF-α), interleukin (IL-1), and Targeting both IL-12 and IL-23, and more recently IL-17 and IL-23 alone , which has led to effective biological treatment (Phase 1 and 2 clinical trials using guselkumab Guselkumab (also known as CNTO1959) inhibits the p19 IL-2 binds to the subunit and is required for terminal differentiation of T helper (Th) 17 cells A fully human IgG1 lambda monoclonal antibody that inhibits the intracellular and downstream signaling of 3 It is. Summary of the Invention [Means for solving the problem]

[0009] In a first aspect, the present invention provides an anti-IL-23 specific antibody (also referred to as IL-23p19 antibody). For example, a method for treating psoriasis in a patient comprising subcutaneously administering guselkumab to the patient. Regarding the anti-IL-23 specific antibody, the first dose, 4 weeks after, and every 8 weeks thereafter It is administered at one dosing interval, for example, at weeks 0, 4, 8, 16, 24, 3, 4, 5, 6, 7, 8, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, Dosing will be done at weeks 2, 40, and 48.

[0010] In a method for treating psoriasis in a patient, the patient is treated with an antibody against IL-23, Clinical endpoints for psoriasis included the antibody secukinumab (Cosen Clinical improvement of psoriasis achieved by patients treated with tyx® (sold under the trademark The efficacy of the drug was higher than that of the endpoints. The clinical endpoints of psoriasis were: PASI90, PASI100, IGA0 and / or IGA1, initial treatment 24 weeks, 28 weeks, 32 weeks, 36 weeks, 40 weeks, 44 weeks, and / or or after 48 weeks, preferably 48 weeks after the initial treatment.

[0011] In the method of the present invention, the antibody against IL-23 is administered at the first administration, 4 weeks after the first administration, and After 4 weeks, the drug is administered every 8 weeks. Secukinumab antibody is administered at the first dose, 1 week after the first dose, 2 weeks after the first dose, 3 weeks after the first dose, 4 weeks after the first dose, and 4 weeks after the first dose. In one embodiment of the method, the antibody is administered every four weeks after the first dose. The antibody against IL-23 was administered at a dose of 100 mg, and the antibody against IL-23 was administered to the scalp, nails, and skin of the patient. It is a safe and effective treatment for psoriasis in areas selected from the group consisting of the hands and feet.

[0012] In another embodiment of the method, an antibody against IL-23 reduces symptoms of psoriasis in a patient. Inducing a clinical response in a patient; Inducing or maintaining clinical remission. , effective for inhibiting disease progression or inhibiting disease complications, Patients with moderate to severe psoriasis are treated.

[0013] In a further embodiment of the invention, the method comprises administering to the patient at least one prior administration of secukinumab. deciding to discontinue treatment of the patient treated with guselkumab and to treat the patient with guselkumab. In further embodiments, the method further comprises administering to the patient 24 weeks, 28 weeks, 32 weeks after the initial treatment. Psoriasis clinical endpoints were assessed at 36, 40, 44 and / or 48 weeks. measuring the PASI90, PASI100, IGA0, and / or IGA1; discontinuing treatment of a patient previously treated with at least one dose of secukinumab. and treating the patient with guselkumab.

[0014] In another aspect, the composition used in the method of the present invention comprises a pharmaceutical composition, The substance is from about 1.0 μg / mL to about 1000 mg / mL, particularly 50 mg / mL or 100 mg In a preferred embodiment, the anti-IL-23 specific antibody is The target antibody was 100 mg / mL guselkumab and 7.9% (w / v) scaffolding of the pharmaceutical composition. 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80 of the medical composition, and the diluent is a standard It is water in a state.

[0015] In one embodiment, the psoriasis patient has cured or minimal disease at 16 weeks. The IGA score (IGA0 / 1) and PASI response corresponding to the above 90% improvement in PASI response (PASI90) or 100% improvement in PASI response (PA The endpoint of achieving SI100 was achieved.

[0016] In another aspect of the invention, the pharmaceutical composition comprises: (i) a peptide comprising a sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:20, and SEQ ID NO: (ii) the heavy chain CDR amino acid sequence of SEQ ID NO:50, SEQ ID NO:56, and SEQ ID NO: and the light chain CDR amino acid sequence of No. 73. 100 mg / mL of anti-IL23 specific antibody and 7.9% (w / v) of the pharmaceutical composition 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, 0.053% (w / v) polysorbate 80 in the pharmaceutical composition; and the diluent is standard It is water in a state.

[0017] Another aspect of the method of the present invention includes administering a pharmaceutical composition, the pharmaceutical composition comprising SEQ ID NO: The guselkumab heavy chain variable region amino acid sequence of SEQ ID NO: 106 and the guselkumab light chain variable region amino acid sequence of SEQ ID NO: 116 and an isolated anti-IL-23 specific antibody having the amino acid sequence of The pharmaceutical composition contained 7.9% (w / v) sucrose and 4.0 mM histidine. , 6.9 mM L-histidine monohydrochloride monohydrate, and 0.053% (w / v) of the pharmaceutical composition. ) polysorbate 80, and the diluent is water at standard conditions. [Brief description of the drawings]

[0018] [Figure 1] The percentage of subjects achieving a PASI90 response, a PASI100 response, an IGA score of cure (0), and an IGA score of cure (0) or minimal (1) from Week 1 through Week 48 are shown. [Diagram 2] FIG. 1 is a diagram of the Eclipse study design. [Diagram 3] 1 shows serum concentrations of IL-17F in psoriasis patients treated with guselkumab and secukinumab. [Figure 4] FIG. 1 shows serum concentrations of IL-22 in psoriasis patients treated with guselkumab and secukinumab. [Diagram 5] 1 shows serum concentrations of BD-2 in psoriasis patients treated with guselkumab and secukinumab. [Figure 6] Normalization of a subset of induced genes in psoriatic lesional skin is shown. [Figure 7] 1 shows the expression of genes related to mucosa-associated invariant T (MAIT) cells in psoriatic lesion skin. [Figure 8] FIG. 1 shows the frequency of CD8-positive tissue-resident memory T cells (TRM) in psoriatic skin treated with guselkumab and secukinumab. [Figure 9] 1 shows the frequency of regulatory T cells (Tregs) in psoriatic skin treated with guselkumab and secukinumab. [Figure 10] Figure 2 shows the ratio of regulatory T cells (Treg) to CD8-positive tissue-resident memory T cells (TRM) in psoriatic skin treated with guselkumab and secukinumab.

[0019] As used herein, the method of treating psoriasis includes administering isolated, recombinant and / or synthetic anti-I L-23 specific human antibodies and diagnostic and therapeutic compositions, methods thereof and and an apparatus for

[0020] As used herein, "anti-IL-23 specific antibody," "anti-IL-23 antibody," "anti The term "antibody fragment" and / or "antibody variant" are incorporated into the antibody of the present invention. At least one complementarity determining region (CDR) or a complementarity determining region (CDR) of the heavy or light chain that can be embedded in the or a ligand-binding portion thereof, a heavy or light chain variable region, a heavy or light chain constant region, A framework region, or any portion thereof, or an IL-23 receptor or binding protein A small amount of an immunoglobulin molecule, such as, but not limited to, at least a portion of a protein. Such antibodies include any protein or peptide-containing molecule that contains at least a portion of the Optionally, further specific ligands may be affected, including but not limited to, Such antibodies can be used to bind to at least one IL-1 in vitro, in situ, and / or in vivo. modulate, reduce, or increase IL-23 activity or binding, or IL-23 receptor activity or binding Non-limiting examples include: Thus, preferred anti-IL-23 antibodies, specified portions or variants of the invention comprise at least one A compound capable of binding to the IL-23 molecule or a specified portion, variant, or domain thereof. A suitable anti-IL-23 antibody, specified portion, or variant may also optionally be At least one IL-23 activity or function may be affected, but such activity or function may not be affected. The properties or functions of the protein include synthesis of RNA, DNA, or protein, release of IL-23, IL-23 receptor signaling, cleavage of membrane IL-23, activity of IL-23, IL-2 These include, but are not limited to, the production and / or synthesis of 3.

[0021] The term "antibody" further includes antibody mimetics, such as single chain antibodies and fragments thereof. Parts of antibodies that mimic the structure and / or function of such antibodies or their specific flags The present invention includes antibodies, digested fragments, specified portions, and variants thereof, including fragments or portions thereof. The functional fragment is intended to include an antigen-binding fragment that binds to mammalian IL-23. For example, Fab (e.g., by papain digestion), Fab' (e.g., by peptide digestion), by pepsin digestion and partial reduction), and F(ab')2 (by, for example, pepsin digestion). , facb (e.g., by plasmin digestion), pFc' (e.g., by pepsin or plasmin digestion), (e.g., by pepsin digestion, partial reduction, and reassembly), F Examples of such fragments include, but are not limited to, scFv or scFv (e.g., by molecular biology techniques) fragments. Antibody fragments capable of binding to IL-23 or portions thereof that are not limited to IL-23 are encompassed by the invention (e.g., (see Colligan, Immunology, supra).

[0022] Such fragments may be used as known in the art and / or as described herein. The antibody can be produced by enzymatic cleavage, synthetic or recombinant techniques, as described in The present invention also provides methods for producing antibody genes in which one or more stop codons have been introduced upstream of the natural stop site. For example, F(ab')2 heavy chain portions can be produced in various truncated forms using The combination of genes that are encoded is H Encoding one domain and / or hinge region The various portions of an antibody can be engineered to contain DNA sequences that encode the antibody. It is possible to chemically bind the protein or use genetic engineering techniques to create a continuous protein sequence. It can be prepared as a protein.

[0023] As used herein, the term "human antibody" refers to an antibody in which substantially all portions of the protein (e.g., For example, CDR, framework, C L , C H Domain (e.g., C H 1. C H 2. C H 3 ), Hinge (V L , V H )) is substantially non-human in nature with only minor sequence changes or mutations. "Human antibody" refers to an antibody that is immunogenic. "Human antibody" also refers to a human germline immunoglobulin. Human antibodies may be antibodies derived from the sequence of the human genome or may be antibodies that match exactly. Amino acid residues not encoded by cell-type immunoglobulin sequences (e.g., in vitro By the introduction of random or site-specific mutations in the target gene or by somatic mutations in vivo In many cases, this is because a human antibody may contain mutations that are specific to the human antibody. Human antibodies are characterized by the similarity of their amino acid sequences, which means that they are substantially non-immunogenic. Therefore, sequence similarity searches can be used to identify similar Selecting an antibody having the linear sequence of as a template for generating a human antibody. Similarly, names can include primates (monkeys, baboons, chimpanzees, etc.), rodents (mouse, Antibodies from such species, including rats, rabbits, guinea pigs, hamsters, etc., and other mammals, It is understood that chimeric antibodies are subgenus, genus, subfamily, and family specific antibodies. Such changes or mutations may optionally and preferably include modified The antibody has the ability to retain or reduce immunogenicity in humans or other species compared to unmodified antibodies. Thus, a human antibody is distinct from a chimeric antibody or a humanized antibody.

[0024] A human antibody is a functionally rearranged human immunoglobulin (e.g., a heavy and / or light chain ) produced by a non-human animal, or a prokaryotic or eukaryotic cell, capable of expressing a gene Moreover, when the human antibody is a single chain antibody, it is possible to obtain a single chain antibody that is similar to a natural human antibody. For example, Fv may contain a linker peptide not found in the heavy chain variable region and a light chain variable region. A linker peptide such as 2 to about 8 glycine or other amino acid residues that connects the variable region Such linker peptides are considered to be of human origin.

[0025] Also, preferred are monoclonal antibodies having binding specificities for at least two different antigens. Bispecific antibodies, heterospecific antibodies, heterologous antibodies, which are preferably human or humanized antibodies. Alternatively, an antibody or a similar antibody may be used, in which case one of the binding specificities is at least One against the IL-23 protein and the other against any other antigen. Methods for producing bispecific antibodies are known in the art. The recombinant production of target antibodies is based on the coexpression of two immunoglobulin heavy-light chain pairs, The two heavy chains have different specificities (Milstein and Cuello, Na ture, 305:537 (1983). Random combination of immunoglobulin heavy and light chains. To combine these hybridomas (quadromas), ten different antibody molecules were produced. of these only one has the correct bispecific structure. Purification of the molecule, usually achieved by affinity chromatography steps, is quite tedious. and the product yield is low. A similar procedure is described, for example, in WO 93 / 08829. , U.S. Patent Nos. 6,210,668, 6,193,967, and 6,132,99 No. 2, No. 6,106,833, No. 6,060,285, No. 6,037,453 No. 6,010,902, No. 5,989,530, No. 5,959,084 , No. 5,959,083, No. 5,932,448, No. 5,833,985, Same No. 5,821,333, Same No. 5,807,706, Same No. 5,643,759, Same No. No. 5,601,819, No. 5,582,996, No. 5,496,549, No. 5,496,549, No. No. 4,676,980, WO 91 / 00360, WO 92 / 00373 , European Patent No. 03089, Traunecker et al., EMB ○J.10 :3655(1991), Suresh et al., Methods in Enz and in J. Med. 121:210 (1986), each of which is incorporated herein by reference. 6,311,421, all of which are incorporated herein by reference in their entirety.

[0026] Anti-IL-23 specific antibodies (IL-23 specific) that are useful in the methods and compositions of the invention Antibodies against IL-23 (also referred to as specific antibodies) have high affinity binding to IL-23. and optionally and preferably, having low toxicity. Specifically, each component, such as the variable region, the constant region, and the framework, is individually and / or collectively, optionally and preferably, the compositions of the present invention have low immunogenicity. The antibodies, or specified fragments or variants thereof, are useful in the present invention. Antibodies that can be used in the present invention have the potential to provide measurable relief of symptoms and low and / or tolerable Optionally, it may be characterized based on its ability to treat patients for long periods of time with minimal toxicity. or acceptable immunogenicity and / or high affinity, as well as other favorable properties, are obtained. "Low immunogenicity" as used herein can contribute to a therapeutic outcome that is favorable for the patient being treated. Less than about 75%, or preferably less than about 50%, of the subjects had significant HAHA, HACA or other increased HAMA response and / or low titers (double antigen enzyme) in treated patients an increase of less than about 300, preferably less than about 100, as measured by immunoassay (Elliott et al., Lancet 344:1125- 1127 (1994), which is incorporated herein by reference in its entirety. "Low Immunogenicity" is observed in less than 25% of patients treated at the recommended dose for the recommended course of therapy during the treatment period, preferably in patients treated with anti-IL-23 antibodies, when It can also be defined as the incidence of titer levels of antibodies to anti-IL-23 antibodies in .

[0027] The terms "clinically proven effective" and "clinically proven effective" refer to As used herein in the context of a dosage regimen, treatment or method, a particular dose, administration Efficacy refers to the progression of a disease in response to the agent of the invention. For example, the anti-IL-23 antibody of the present invention (e.g., anti-IL-23 antibody) can be used to measure the change in the expression level of IL-23. -23 antibody guselkumab) is a marker that reflects the severity of the disorder being treated. and administering to the patient in an amount and for a time sufficient to cause improvement, preferably sustained improvement, in To determine whether the amount and duration of treatment is sufficient, Various indicators that reflect the extent of the disease, disorder, or condition can be assessed. Such indicators include, for example: Clinically recognized risk of disease severity, symptoms, or manifestations of the disorder of interest. The degree of improvement is assessed overall by the physician, who reports this assessment on the basis of symptoms, It can be based on symptoms, biopsy, or other test results, and can also be conducted by questionnaires given to subjects. Use a questionnaire, such as a quality of life questionnaire developed for a given disease. For example, the anti-IL-23 antibodies of the invention may be used to improve a patient's condition associated with psoriasis. This improvement can be achieved by improving disease activity indicators, remission of clinical symptoms, or any other measure of disease activity. Examples are PASI75, PASI90, PASI100, IGA1, and IGA0. The Psoriasis Area and Severity Index (PASI) is a tool used by doctors and nurses to record the severity of psoriasis. The PASI score used by the PASI Centers for Clinical Neurology, PASI 75 is the PASI score from start to finish. It is an abbreviated expression for a 75% reduction in score (note that PASI90 means a 90% reduction). PASI100 means 100% reduction). Investigator Global Assessment (IGA The IGA tool is a visual assessment consisting of a score ranging from 0 (cured) to 4 (severe). 0 means cured and IGA1 means almost cured.

[0028] The term "having clinically proven safety" refers to an anti-IL-23 antibody of the present invention (e.g. For example, the dosage, drug administration method, treatment, or method of using the anti-IL-23 antibody guselkumab If the drug is approved, there is a favorable risk:benefit ratio compared with standard of care or another comparator drug, and no treatment-emergent events are expected to occur. The frequency of adverse events (termed AEs or TEAEs) occurring is acceptable and / or severe. Adverse events are events that occur in patients who have been administered a drug. This is an undesirable medical occurrence. In particular, the dosage and administration of the anti-IL-23 antibodies of the present invention Regimen- or treatment-related safety is uncertain, as adverse events may be due to the use of anti-IL-23 antibodies. Adverse events associated with antibody administration if considered likely, probable, or highly probable This refers to the acceptable frequency and / or acceptable severity of the disorder.

[0029] As used herein, unless otherwise specified, the term "clinically proven" or to modify the terms "safety" and / or "effectiveness" FDA, EMEA, or equivalent national regulatory agency approval criteria. For example, a clinical trial is a test to determine whether a drug has been approved for use in a clinical trial. An adequately sized randomized, double-blind study used to clinically prove efficacy. This is also possible.

[0030] usefulness The isolated nucleic acids of the present invention comprise at least one anti-IL-23 antibody or a specific variant thereof. The invention can be used to produce a compound that can diagnose, monitor, regulate, treat, or alleviate the symptoms of psoriasis. , to help prevent or reduce the occurrence of symptoms of psoriasis, (including mammals and humans) can be done.

[0031] Such methods include those for such regulating, treating, alleviating, preventing, or reducing symptoms, or for treating or preventing a disease. The method comprises administering to a cell, tissue, organ, animal or patient in need thereof at least one anti-IL- The method may include administering an effective amount of a composition or pharmaceutical composition comprising antibody 23. This can be done using known methods, as described herein or as known in the relevant art. When determining the dosage, it is about 0.001 per single (e.g., bolus), multiple, or continuous administration. ~500mg / kg or 0.01-500 per single, multiple, or continuous dose 0 μg / mL, or any effective range or value therein. That's fine too.

[0032] References All publications or patents cited herein, whether specifically stated or not, are hereby incorporated by reference. which are incorporated herein in their entirety and represent prior art at the time of the present invention; and / or A publication is a document recorded in electronic or printed form that describes and enables the invention. Any scientific publication or patent available in any media format, including any The following documents are incorporated herein by reference in their entirety: :Ausubel,et al.,ed.,Current Protocols i n Molecular Biology,John Wiley&Sons,Inc. ,NY,NY(1987-2001),Sambrook,et al.,Molecu lar Cloning:A Laboratory Manual,2nd Edit ion, Cold Spring Harbor, NY (1989), Harlow a nd Lane,antibodies,a Laboratory Manual,C old Spring Harbor, NY (1989), Colligan, et a l.,eds.,Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001), Colli gan et al.,Current Protocols in Protein Science,John Wiley&Sons,NY,NY,(1997-2001 ).

[0033] Antibodies of the invention - production and purification The at least one anti-IL-23 used in the methods of the invention is optionally a compound known in the art. Any cell line, mixed cell line, immortalized cell, or clonal population of immortalized cells known in the art. For example, Ausubel, et al., ed., Cur rent Protocols in Molecular Biology,John Wiley & Sons, Inc., NY, NY (1987-2001), Sambro ok,et al.,Molecular Cloning:A Laboratory Manual,2nd Edition,Cold Spring Harbor,N Y (1989), Harlow and Lane, antibodies, a Lab. oratory Manual,Cold Spring Harbor,NY(198 9), Colligan,et al.,eds.,Current Protocol s in Immunology,John Wiley&Sons,Inc.,NY( 1994-2001), Colligan et al.,Current Proto cols in Protein Science,John Wiley&Sons, NY, NY, (1997-2001), each of which is incorporated herein by reference in its entirety. is incorporated into the book.

[0034] A preferred anti-IL-23 antibody has the heavy chain variable region amino acid sequence of SEQ ID NO: 106 and the heavy chain variable region amino acid sequence of SEQ ID NO: and the light chain variable region amino acid sequence of SEQ ID NO:5, SEQ ID NO:20, and SEQ ID NO:116. The heavy chain CDR amino acid sequence of sequence number 44 and SEQ ID NO:50, SEQ ID NO:56, and SEQ ID NO:7 and guselkumab (also referred to as CNTO1959), having the light chain CDR amino acid sequence of Other anti-IL-23 antibodies have the sequences listed herein, the entireties of which are incorporated by reference. No. 7,935,344, which is incorporated herein by reference.

[0035] Human antibodies specific for human IL-23 protein or fragments thereof are isolated IL-23 Suitable molecules such as proteins and / or portions thereof (including synthetic molecules such as synthetic peptides) Other specific or general mammalian antibodies may be raised against the immunogenic antigen. Preparation of immunogenic antigens and generation of monoclonal antibodies may be accomplished by any suitable technique. It can be done using.

[0036] In one approach, a suitable immortal cell line (e.g., but not limited to, Sp2 / 0, S p2 / 0-AG14, NSO, NS1, NS2, AE-1, L.5, L243, P3X6 3Ag8.653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 SA5 , U937, MLA 144, ACT IV, MOLT4, DA-1, JURKAT, W EHI, K-562, COS, RAJI, NIH 3T3, HL-60, MLA 144 Myeloma cell lines such as NAMALWA, NEUR ○2A, or heteromyelomas, The fusion product, or any cell or fusion cell derived therefrom, or any cell or fusion cell derived therefrom, Any other suitable cell line known in the art) (see, for example, www.atcc.org, www.lif etech.com), including, but not limited to, isolated or cloned Antibody cells such as cultured spleen, peripheral blood, lymph, tonsils, or other immune or B cell-containing cells. The nucleic acid may be derived from recombinant or endogenous, viral, cellular, or endogenous cells, or from endogenous or heterologous nucleic acids. Fungi, algae, prokaryotes, amphibians, insects, reptiles, fish, mammals, rodents, horses, sheep, goats , sheep, primates, eukaryotes, genomic DNA, cDNA, rDNA, mitochondrial DNA A or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single As single-, double- or triple-stranded, hybridized, etc., or any combination thereof Either the heavy or light chain constant or variable, or framework or CDR sequences Hybridomas are produced by fusing the cells with any other cells expressing the above vector. See Ausubel, supra, and Colligan, Immunology, chapter 2, supra. both of which are incorporated herein by reference in their entireties.

[0037] Antibody producing cells can also be isolated from the peripheral blood of humans or other suitable animals immunized with an antigen of interest, or Preferably, the host cell may be obtained from the spleen or lymph nodes. Any other suitable host cell may be used. and the like, to produce a heterologous nucleic acid encoding an antibody, specified fragment, or variant thereof of the present invention. It is also possible to express the nucleic acid or endogenous nucleic acid. The cells may be isolated using selective culture conditions or other suitable known methods and isolated by limiting dilution or The desired specificity can be obtained by cloning using cell sorting or other known methods. Cells producing antibodies that bind to the antibody can be selected by a suitable assay (e.g., ELISA). can be done.

[0038] Selecting recombinant antibodies from a peptide or protein library (e.g., bacteriophage Display live phage, ribosomes, oligonucleotides, RNA, cDNA, etc. Examples of antibodies include, but are not limited to, Cambridge antibody T echnologies,Cambridgeshire,UK,MorphoSys, Martinsreid / Planegg, DE, Biovation, Aberdee n,Scotland,UK,BioInvent,Lund,Sweden,Dyax Corp., Enzon, Affymax / Biosite, Xoma, Berkel ey,CA, Ixsys. For example, European Patent No. 368,684, International Application PCT / GB 91 / 01134, International Application No. PCT / GB92 / 01755, International Application No. PCT / GB 92 / 002240, International Application No. PCT / GB92 / 00883, International Application No. PCT / G B93 / 00605, U.S. Patent Application No. 08 / 350260 (5 / 12 / 94), International Application No. PCT / GB94 / 01422, International Application No. PCT / GB94 / 02662, International Application No. PCT / GB97 / 01835, (CAT / MRC), International Publication No. 90 / 1444 No. 3 International Publication No. 90 / 14424 International Publication No. 90 / 14430 International Application PCT / US 94 / 1234, WO 92 / 18619, WO 96 / 07754 (Sc ripps), WO 96 / 13583, WO 97 / 08320 (Mor phoSys), International Publication No. 95 / 16027 (BioInvent), International Publication No. No. 8 / 06630, WO 90 / 3809 (Dyax), U.S. Pat. No. 692 (Enzon), International Application No. PCT / US91 / 02989 (Affymax) , International Publication No. 89 / 06283, European Patent No. 371998, European Patent No. 550400 No., (Xoma), European Patent No. 229046, International Application PCT / US91 / 07149 No. (Ixsys), or stochastically generated peptides or proteins - U.S. Pat. No. 5723323, No. 5763192, No. 5814476, No. 5817483 No. 5824514, No. 5976862, International Publication No. 86 / 05803, State Patent No. 590,689 (Ixsys, Applied Molecular Evolution) (AME) and its predecessors, each of which is incorporated herein by reference in its entirety. The present invention relates to a method for the preparation of ... compound having a molecular weight of 1,000 or more, which is incorporated herein by reference, or which is known in the art and / or described herein. The present invention relates to a method for producing a repertoire of human antibodies in a transgenic animal. It depends on the immunization (e.g., SCID mice, Nguyen et al., Microb iol.Immunol.41:901-907(1997), Sandhu et a l., Crit. Rev. Biotechnol. 16:95-118 (1996), E Ren et al., Immunol. 93:154-161 (1998) (which are incorporated by reference in their entireties, and related patents and applications) including, but not limited to, methods Other suitable methods for generating or isolating antibodies of the required specificity may be used, including but not limited to: Such techniques include ribosome display (Hanes et al., Proc. c.Natl.Acad.Sci.USA,94:4937-4942(May 199 7), Hanes et al., Proc. Natl. Acad. Sci. USA, 9 5:14130-14135 (Nov. 1998)), single cell antibody generation technology (e.g., Selected Lymphocyte Antibody Method ("SLAM") (U.S. Pat. No. 5,627,052, Wen et al. t al., J. Immunol. 17:887-892 (1987), Babcook et al.,Proc.Natl.Acad.Sci.USA 93:7843-7 848 (1996)), gel microdroplets and flow cytometry (Pow ell et al.,Biotechnol.8:333-337(1990), On e 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 selection (St eenbakkers et al.,Molec.Biol.Reports 19: 125-134(1994), Jonak et al., Progress Biot ech,Vol.5,In Vitr ○Immunization in Hybri doma Technology,Borrebaeck,ed.,Elsevier Science Publishers BV,Amsterdam,Nether Lands (1988) are examples of such compounds, but are not limited to these.

[0039] Methods for engineering or humanizing non-human or human antibodies can be used as well. Generally, humanized or engineered antibodies are antibodies that are produced by a non-human, e.g. including, but not limited to, from mouse, rat, rabbit, non-human primate, or other mammal These non-human amino acid residues are often referred to as " In the present case, the amino acid sequence is replaced by a residue called an "import" residue. Such an "import" residue is Typically, "import" variable, constant or other domains of a known human sequence. Obtained from the in.

[0040] Known human Ig sequences are disclosed, for example, at: www.ncbi.nlm.nih.gov / IgSequences. ih.gov / entrez / query.fcgi, www.ncbi.nih.go v / igblast、www.atcc.org / phage / hdb.html、ww w.mrc-cpe.cam.ac.uk / ALIGNMENTS.php、www.k abatdatabase.com / top.html、ftp.ncbi.nih.g ov / repository / kabat、www.sciquest.com、www .abcam.com、www.antibodyresource.com / onli necomp.html、www.public.iastate.edu / ~pedr o / research_tools.html、www.whfreeman.com / immunology / CH05 / kuby05.htm、www.hhmi.org / grants / lectures / 1996 / vlab、www.path.cam.a c.uk / ~mrc7 / mikeimages.html、mcb.harvard.e du / BioLinks / Immunology.html、www.immunolo gylink.com、pathbox.wustl.edu / ~hcenter / in dex.html、www.appliedbiosystems.com、www.n al.usda.gov / awic / pubs / antibody、www.m.ehi me-u.ac.jp / ~yasuhito / Elisa.html、www.biod esign.com、www.cancerresearchuk.org、www.b iotech.ufl.edu、www.isac-net.org、baserv.u ci.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.ht ml, http: / / www.bioinf.org.uk / abs, antibody .bath.ac.uk, www.unizh.ch, www.cryst.bbk.a c.uk / ~ubcg07s, www.nimr.mrc.ac.uk / CC / ccae wg / ccaewg.html, www.path.cam.ac.uk / ~mrc7 / humanisation / TAHHP.html, www.ibt.unam.mx / vir / structure / stat_aim.html, www.biosci.m issouri.edu / smithgp / index.html, www.jerin i.de, Kabat et al., Sequences of Proteins of Immunological Interest,USDept.Healt h (1983), each of which is incorporated herein by reference in its entirety.

[0041] Such imported sequences may be used to reduce immunogenicity or to enhance the expression of polypeptides, as described in the art. As known in the art, the term "binding," "affinity," "association rate constant," "dissociation rate constant," "binding activity," "specificity," The compounds may be used to reduce, enhance or modify the half-life, half-life, or any other suitable property. Generally, the CDR residues directly and most substantially influence antigen binding. Thus, it is possible to maintain some or all of the non-human or human CDR sequences while modifying the CDRs with variable The non-human sequences in the regions and constant regions can be replaced with human amino acids or other amino acids. It is also possible.

[0042] Antibodies may optionally be humanized, or human antibodies have high affinity for the antigen. To this end, the genetic material may be modified while retaining its sex and other advantageous biological properties. Optionally, a three-dimensional model of the parental and humanized sequences is used to identify the parental and various Humanized (or human) antibodies can be prepared by a process of analyzing theoretical humanized products. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. The most likely three-dimensional structures of selected immunoglobulin sequence candidates have been determined. Computer programs are available for illustrating and displaying the images. The analysis of the likely role of residues in the function of the candidate immunoglobulin sequence. In other words, it is possible to analyze residues that affect the antigen-binding ability of immunoglobulin candidates. As a result, desirable antibody properties such as enhanced affinity for the target antigen(s) are achieved. Framework (FR) residues were selected from the consensus and import sequences so that You can choose and combine them.

[0043] In addition, the human IL-23 specific antibodies used in the methods of the present invention contain human germline light chain fragments. In certain embodiments, the light chain germline sequence may include the following: A1, A10, A 11, 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 / 18 a, L5, L6, L8, L9, O1, O11, O12, O14, O18, O2, O4, and The VK sequences are selected from human VK sequences, including, but not limited to, O8 and O9. In this embodiment, the light chain human germline frameworks are V1-11, V1-13, V1-16, V 1-17, V1-18, V1-19, V1-2, V1-20, V1-22, V1-3, V 1-4, V1-5, V1-7, V1-9, V2-1, V2-11, V2-13, V2-1 4, 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, Selected from V5-2, V5-4, and V5-6.

[0044] In another embodiment, the human IL-23 specific antibody used in the methods of the invention is a human genotype. In certain embodiments, the heavy chain human germline framework may comprise a human germline heavy chain framework. Frameworks are 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, VH 3-38, 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.

[0045] 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., two or three, such as FR2 and FR3) In certain embodiments, the nucleic acid sequence includes at least FRL1, FRL2, F In another embodiment, at least FRH1, FRL3, or FRL4 are fully human. RH2, FRH3, or FRH4 is fully human. Each of FRL1, FRL2, FRL3, and FRL4 is a germline sequence (e.g., a human germline sequence). sequence), or a human consensus sequence for a particular framework (see above). In another embodiment, the human Ig sequence comprises at least FRH1, FRH2, FRH3, or FRH4 may be selected from the group consisting of germline sequences (e.g., human germline sequences). sequence) or comprises a human consensus sequence for a particular framework. In an embodiment, the framework regions are completely human framework regions.

[0046] The humanization or engineering of the antibodies of the present invention can be carried out according to the methods described in Winter (Jones et al., Nature 321:522(1986), Riechmann et al.,Na ture 332:323 (1988), Verhoeyen et al. nce 239:1534(1988)), Sims et al., J. Immuno l.151:2296(1993), Chothia and Lesk, J. Mol. Biol.196:901(1987), Carter et al., Proc.Na tl.Acad.SCi.USA89:4285(1992), Presta e t al., J. Immunol. 151:2623 (1993), U.S. Patent No. 5723 No. 323, No. 5976862, No. 5824514, No. 5817483, No. No. 5814476, No. 5763192, No. 5723323, No. 5,76688 No. 6, No. 5714352, No. 6204023, No. 6180370, No. 56 No. 93762, No. 5530101, No. 5585089, No. 5225539, No. 4,816,567, International Application PCT / :US98 / 16280, US96 / 189 No. 78, US91 / 09630, US91 / 05939, US94 / 01234, No. GB89 / 01334, GB91 / 01134, GB92 / 01755, International Publication No. 90 / 14443, WO 90 / 14424, WO 90 / 14430 and European Patent No. 229246 (each of which is incorporated herein by reference in its entirety). Any known method, including but not limited to those described in the literature (including those described in the literature used in the literature) may be used. This can be done using the method of

[0047] In some embodiments, the antibody comprises an altered (e.g., mutated) Fc region. For example, in some embodiments, the Fc region provides effector functions of the antibody. In some embodiments, the Fc region is modified to reduce or enhance the I The isotype is selected from IgM, IgA, IgG, IgE, or other isotypes. Alternatively or in addition, amino acid modifications and C1q binding of the Fc region of the IL-23 binding molecule may be used. and / or one or more additional amino acid modifications that alter complement-dependent cytotoxicity function. Starting polypeptides of particular interest are those that bind C1q. It may be a compound that exhibits complement-dependent cytotoxicity (CDC). Polypeptides having the ability to mediate CDC and, optionally, to mediate these activities The antibody may be modified to enhance one or both of the following properties: and / or the amino acid modifications that modify the complement-dependent cytotoxicity function thereof are described, for example, in WO 02 / 06366. No. 42072, incorporated herein by reference.

[0048] As disclosed above, for example, C1q binding and / or FcγR binding may be modified, Complement-dependent cytotoxicity (CDC) activity and / or antibody-dependent cell-mediated cytotoxicity ( By altering antibody-dependent cell-mediated cytotoxicity (ADCC) activity, Thus, the Fc of the human IL-23 specific antibody of the present invention having altered effector functions An "effector function" can be designed to affect the function of a living organism (e.g., in a subject). Examples of effector functions include, but are not limited to, those that activate or reduce the activity of a protein. Although not widely used, C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, and cytotoxicity are These include downregulation of cell surface receptors (e.g., B cell receptor, BCR). Such effector functions are achieved by the Fc region acting as a binding domain (e.g., an antibody variable domain). A variety of assays (e.g., Fc binding assays) may be required. The antibody can be evaluated using various assays (e.g., ELISA, ADCC assay, CDC assay, etc.).

[0049] For example, having improved C1q binding and improved FcγRIII binding (e.g., Human IL-23 (also known as IL-23A) with both improved ADCC activity and improved CDC activity Alternatively, the effector When it is desired to reduce or eliminate a function, variants of the Fc region may be used that inhibit CDC activity. and / or to reduce ADCC activity. In embodiments, only one of these activities may be enhanced, optionally simultaneously. Other activities may be reduced (e.g., improved ADCC activity and reduced CDC activity). and vice versa).

[0050] Fc mutations alter interactions with fetal Fc receptors (FcRn) and their pharmacokinetics Genes can also be engineered and introduced to improve properties. A collection of improved human Fc variants has been described (Shields et al., 2002). 001).High resolution mapping of the bind ing site on human IgG1 for FcγRI,FcγRII, FcγRIII,and FcRn and design of IgG1 vari ants with improved binding t ○the FCγR, J .Biol.Chem.276:6591-6604).

[0051] Another type of amino acid substitution is to modify the glycosylation paclitaxel of the Fc region of a human IL-23 specific antibody. Glycosylation of the Fc region is typically N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. O-linked glycosylation can also occur with 5-hydroxyproline or 5-hydroxylysine. Hydroxyamino acids, most commonly serine or threonine, may be used. one of the following sugars: N-acetylgalactosamine, galactose, or xylose Attachment refers to the recognition of an asparagine side chain for the enzymatic addition of a carbohydrate moiety to a peptide sequence. The sequences are asparagine-X-serine and asparagine-X-threonine, where X is proline. Any amino acid other than phosphate. Therefore, it is possible to have any of these peptides in a polypeptide. The presence of this tide sequence provides a site where glycosylation can occur.

[0052] A glycosylation pattern can refer to, for example, one or more glycosylations found on a polypeptide. Deleting a site(s) and / or removing one or more groups that are not present in the polypeptide The Fc of the human IL-23 specific antibody can be modified by adding glycosylation sites. The addition of glycosylation sites to the region can be accomplished by modifying the region to include one or more of the above tripeptide sequences. This is successfully achieved by modifying the amino acid sequence (in the case of N-linked glycosylation sites). An exemplary glycosylation variant has an amino acid substitution at residue Asn297 of the heavy chain. This alteration may involve the addition of one or more serine or threonine residues to the original polypeptide sequence. or by substitution therewith (for O-linked glycosylation sites). In addition, changing Asn 297 to Ala removes one of the glycosylation sites. can be done.

[0053] In certain embodiments, the human IL-23 specific antibodies of the invention bind to GnT III. To add 1cNAc to the human IL-23 antibody, beta(1,4)-N-acetylglucosamine was synthesized. Expressed in cells expressing glucosaminyltransferase III (GnT III) Methods for producing antibodies in such a manner are described in WO 9954342, WO 9954343, WO 995434 ... Publication No. 03011878, Patent Publication No. 20030003097A1, and Umana et al.,Nature Biotechnology,17:176-180, Feb. 1999, all of which are incorporated herein by reference in their entireties. Specifically incorporated.

[0054] Anti-IL-23 antibodies also optionally include those described herein and / or in the art. As is known in the art, transgenic mice capable of producing a repertoire of human antibodies have been developed. Produced by immunization of a control animal (e.g., mouse, rat, hamster, non-human primate, etc.) Cells producing human anti-IL-23 antibodies may be produced as described herein. They may be isolated from such animals and immortalized using any suitable method.

[0055] Transgenic mice capable of generating a repertoire of human antibodies that bind to human antigens The cuminoids can be produced by known methods, including, but not limited to, However, U.S. Patents 5,770,428 and 5,569, issued to Lonberg et al. No. 825, No. 5,545,806, No. 5,625,126, No. 5,625,8 25, 5,633,425, 5,661,016, and 5,789, No. 650, WO 98 / 50433 by Jakobovits et al. ts et al., International Publication No. 98 / 24893; Lonberg et al., International Publication No. 98 / 2488 No. 4, International Publication No. 97 / 13852 by Lonberg et al. No. 94 / 25585, Kucherlapate et al., International Publication No. 96 / 34096, European Patent No. 0463151(B1) to Kucherlapate et al. European Patent No. 0710719(A1) to Pate et al., U.S. Patent No. 5,500,000 to Surani et al. 45,807, Bruggemann et al., International Publication No. 90 / 04036, Brugg Emann et al., European Patent No. 0438474(B1); Lonberg et al., European Patent No. No. 814259(A2), Lonberg et al., UK Patent No. 2272440(A), Lonberg et al. Nature 368:856-859 (1994), T aylor et al.,Int.Immunol.6(4)579-591(199 4), Green et al., Nature Genetics 7:13-21(1 994), Mendez et al., Nature Genetics 15:14 6-156 (1997), Taylor et al., Nucleic Acids Research 20(23):6287-6295(1992), Tuaillon et al.,Proc Natl Acad Sci USA 90(8)3720 -3724(1993), Lonberg et al., Int Rev Immun ol 13(1):65-93(1995), and Fishwald et al., at Biotechnol 14(7):845-851(1996), these are references. (Each of the above references is incorporated herein in its entirety by reference). In general, these mice At least one human immunoglobulin that has been or can undergo functional rearrangement. The mouse comprises at least one transgene comprising DNA derived from the chromosome locus. The endogenous immunoglobulin locus of the mouse is disrupted or deleted to generate the endogenous immunoglobulin gene of the mouse. The ability to produce antibodies encoded by the vector can be eliminated.

[0056] Screening antibodies for specific binding to similar proteins or fragments This can be successfully achieved using peptide display libraries. A large collection of peptides for individual components with a desired function or structure Antibody screening of peptide display libraries. The length of the displayed peptide sequence is 3 to 100 μg / s. More than 5000 amino acids, frequently 5-100 amino acids long, most often about 8-2 In addition to direct chemical synthesis methods for generating peptide libraries, Several recombinant DNA methods have also been described. One type uses bacteriophages. Each bacteriophage or cell includes displaying a peptide sequence on its surface. The cells contain nucleotide sequences that code for specific displayed peptide sequences. Such methods are described in WO 91 / 17271, WO 91 / 18980, WO 91 / 19063 and WO 91 / 19064. No. 19818, and No. 93 / 08278.

[0057] Other systems for generating peptide libraries include in vitro chemical synthesis and recombinant The present invention has both aspects of the method described in International Publication Nos. 92 / 05258 and 92 / 14843. and 96 / 19256. Also see U.S. Pat. Nos. 5,658,754 and See also U.S. Pat. Nos. 5,643,768 and 5,643,768. Peptide display libraries, vectors The screening kit was provided by Invitrogen (Carlsbad, CA) and and Cambridge antibody Technologies (Cambrid They are commercially available from sources such as Enzon (Essex, England). Nos. 4,704,692, 4,939,666, 4,946,778, and No. 5260203, No. 5455030, No. 5518889, No. 553462 No. 1, No. 5656730, No. 5763733, No. 5767260, No. 58 No. 56456, No. 5223409, No. 5403484, and No. 56456, which were assigned to Dyax. Nos. 5,571,698, 5,837,500, and 5,427 assigned to Affymax No. 908, No. 5580717, Cambridge antibody Techn. No. 5,885,793 assigned to Genentech; No. 5750373, No. 5618920 and No. 5595898 assigned to Xoma No. 5576195, No. 5698435, No. 5693493, No. 569 No. 8417, Colligan, above, Ausubel, above, or Sambro, above ok, each of which is incorporated herein by reference in its entirety. To be incorporated.

[0058] The antibodies used in the methods of the present invention may also be derived from goats, cows, horses, and other animals that produce such antibodies in their milk. The nucleic acid can be used to provide a transgenic animal or mammal, such as a mouse, sheep, rabbit, etc. It may also be prepared using at least one anti-IL-23 antibody encoding the same. Animals can be prepared using known methods, including, but not limited to, However, U.S. Patent Nos. 5,827,690, 5,849,992, and 4,873,3 No. 16, No. 5,849,992, No. 5,994,616, No. 5,565,36 2, 5,304,489, etc. (each of which is incorporated by reference in its entirety). is incorporated herein).

[0059] The antibodies used in the method of the present invention may be used in plant parts or cells cultured therefrom. Transgenic plants and cultured plants that produce such antibodies, specified portions, or variants. providing plant cells (e.g., but not limited to, tobacco and corn) The method further comprises using at least one anti-IL-23 antibody-encoding nucleic acid to prepare a As a non-limiting example, a recombinant protein can be expressed using an inducible promoter. Transgenic tobacco leaves expressing β-lactamase were successfully used to provide large amounts of recombinant protein. For example, Cramer et al., Curr. Top. Microbo I. Immunol. 240:95-118(1999) and references cited therein. Transgenic maize has also been produced in other recombinant systems. Proteins that have biological activity equivalent to those produced or purified from natural sources. It has been used to express mammalian proteins at commercial production levels. d et al.,Adv.Exp.Med.Biol.464:127-147(19 99) and the references cited therein. Antibodies may also be single chain antibodies (scFv). Transgenic plants such as tobacco seeds and potato tubers containing antibody fragments such as It has also been produced in large quantities from the seeds of some phytoplankton plants. lant Mol. Biol. 38:101-109 (1998) and references therein. Therefore, the antibodies of the present invention can also be transfected according to known methods. Transgenic plants can also be used for production. For example, Fischer et al. l.,Biotechnol.Appl.Biochem.30:99-108(Oct. .,1999),Ma et al.,Trends Biotechnol.13:5 22-7(1995), Ma et al., Plant Physiol.109:3 41-6(1995), Whitelam et al.,Biochem.Soc.T rans. 22:940-944 (1994), and references cited therein. Each of the above references is incorporated herein by reference in its entirety.

[0060] The antibodies used in the methods of the present invention have a wide range of affinities (K D ) to human IL-23 In a preferred embodiment, the human mAb is optionally capable of binding to a high affinity For example, a human mAb can bind to human IL-23. about 10 -7 M or less, for example, 0.1 to 9.9 (or any range or value therein) x 10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 Or Any range or value in the range, including but not limited to, K D It can be combined with Cut.

[0061] The affinity or avidity of an antibody for an antigen can be determined experimentally using any suitable method. (For example, Berzofsky, et al., "Antibody- Antigen Interactions,” In Fundamental Imm unology, Paul, WE, Ed., Raven Press: New Yo rk, NY (1984), Kuby, Janis Immunology, WHFr Eeman and Company: New York, NY (1992), and Honmei (See methods described in the subsection.) Parenteral antibodies that are measured for specific antibody-antigen interactions Compatibility may differ when measured under different conditions (e.g., salt concentration, pH). Therefore, affinity and other antigen binding parameters (e.g., K D、 K a , K d ) is preferred. or standard solutions of antibodies and antigens, and standard buffers, such as those described herein. This is done using

[0062] nucleic acid molecule Among other sequences disclosed herein, for example, the light chain or At least 70 to 10 contiguous amino acids of at least one of the heavy chain variable or CDR regions 0% of the nucleotide sequence encoding the The present invention also relates to a method for the preparation of a nucleic acid sequence comprising the steps of: The present invention provides a method for the production of a compound according to the present invention, which encodes at least one anti-IL-23 antibody, using the information provided in the present application. The disclosed nucleic acid molecules can be synthesized using methods described herein or known in the art. can be obtained.

[0063] The nucleic acid molecules of the present invention may be in the form of mRNA, hnRNA, tRNA or any other suitable form. in the form of RNA, or cDNA obtained by cloning or produced synthetically and genomic DNA, or any of these forms of DNA. The DNA may be triple-stranded, double-stranded, or single-stranded, or a combination of these. Any combination of any portion of at least one strand of DNA or RNA. The coding strand, also known as the sense strand, may be the coding strand, or may be called the antisense strand. It may also be the non-coding strand.

[0064] The isolated nucleic acid molecule used in the methods of the invention optionally contains one or more introns. An open reading frame (ORF), for example, At least one heavy or light chain CDR1, CDR2, and / or CDR3 is not included. A nucleic acid molecule comprising at least one specified portion of at least one CDR, such as R3. , a nucleic acid molecule comprising a coding sequence of an anti-IL-23 antibody or a variable region, and Although substantially different, due to the degeneracy of the genetic code, the sequences described herein and / or in the art may be used interchangeably. The nucleic acid sequence encoding at least one anti-IL-23 antibody known in the art is also It will be appreciated that the genetic code may include nucleic acid molecules that contain the nucleotide sequence. Therefore, those skilled in the art will be familiar with the specific antibodies to be used in the methods of the present invention. It is routine to generate such modified nucleic acid variants encoding IL-23 antibodies. See, e.g., Ausubel et al., supra, and such nucleic acid variants may be used in accordance with the present invention. Non-limiting examples of isolated nucleic acid molecules include those having HC CDR1 and HC CDR2, respectively. , HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CD Nucleic acids encoding R3 are included.

[0065] As described herein, a nucleic acid molecule comprising a nucleic acid encoding an anti-IL-23 antibody; The antibody fragment itself may encode the amino acid sequence of a full-length antibody or an antibody fragment. a sequence encoding a part of the body, a coding sequence for an antibody, fragment or part, and additional sequences; With or without additional coding sequences as described above, such as at least one intron. without including non-coding 5' and 3' sequences, e.g., splicing and polyadenylation signals. Transcription, mRNA processing, including (e.g., mRNA ribosome binding and stability) Additional non-coding sequences, including but not limited to transcribed, non-translated sequences that play a role in The sequence includes at least one coding sequence for a signal leader or fusion peptide, Additional coding sequences may be included that code for additional amino acids, e.g., amino acids that provide additional functions. The antibody coding sequence can include, but is not limited to, For example, the marker sequence can be fused to an antibody to which it is fused. The polypeptide may be a sequence encoding a peptide that facilitates purification of the antibody containing the polypeptide fragment or portion.

[0066] Polynucleotides that selectively hybridize to the polynucleotides described herein The method of the present invention provides a method for selectively hybridizing to the polynucleotides disclosed herein. The present invention therefore uses isolated nucleic acids that hybridize under hybridization conditions. The polynucleotides of the embodiments are useful for isolating, detecting, and analyzing nucleic acids that contain such polynucleotides. For example, the polynucleotides of the present invention can be used to measure and / or quantify to identify, isolate, or amplify partial or full-length clones in the deposited library. In some embodiments, the polynucleotide is an isolated or otherwise complementary to the cDNA of a human or mammalian nucleic acid library. The sequence may be a genome sequence or a cDNA sequence.

[0067] Preferably, the cDNA library contains at least 80% of the full-length sequences, preferably the full-length It is preferred that the nucleic acid sequence contains at least 85% or 90% of the sequence, more preferably at least 95% of the full-length sequence. This cDNA library can be normalized to increase the representation of rare sequences. Low or medium stringency, using sequences with low sequence identity to the complementary sequence The hybridization conditions are typical, but not limited to, those described above. For higher sequences, medium and high stringency conditions can optionally be used. Low stringency conditions allow selective hybridization of sequences with approximately 70% sequence identity. can be used to identify orthologous or paralogous sequences. Cut.

[0068] Optionally, the polynucleotide encodes at least a portion of an antibody. The nucleotides are capable of selectively hybridizing to the polynucleotides encoding the antibodies of the present invention. For example, the above-mentioned Ausube See Colligan, supra, each of which is incorporated herein by reference in its entirety. Be absorbed.

[0069] Nucleic acid construction Isolated nucleic acids can be prepared by any of a number of methods, including (a) recombinant methods, (b) synthetic methods, as are well known in the art. (c) purification techniques, and / or (d) a combination thereof. can be done.

[0070] The nucleic acid can conveniently include sequences in addition to the polynucleotides of the present invention. For example, a multiple cloning site containing one or more endonuclease restriction sites may be inserted into the nucleobase. The nucleic acid can be inserted into the nucleic acid to aid in the isolation of the polynucleotide. can be inserted to aid in the isolation of the translated polynucleotide of the present invention. For example, a hexahistidine marker sequence provides a convenient means for purifying the proteins of the invention. The nucleic acids of the invention (excluding the coding sequences) are optionally provided as polynucleotides of the invention. Vectors, adapters, or linkers for cloning and / or expression of peptides. .

[0071] Additional sequences may be added to such cloning and / or expression sequences to facilitate cloning and and / or optimizing their function in expression, aiding in the isolation of polynucleotides. It may be possible to improve the transfer of polynucleotides into cells. The use of vectors, adapters, and linkers is well known in the art. (See, e.g., Ausubel, supra, or Sambrook, supra. )

[0072] Recombinant methods for constructing nucleic acids Isolated nucleic acids, such as RNA, cDNA, genomic DNA, or any combination thereof. The acid composition may be derived from a biological source using any number of cloning methods known to those of skill in the art. In some embodiments, the polynucleotides of the present invention can be subjected to stringent amplification. Oligonucleotide probes that selectively hybridize under optimal conditions are prepared by cleaving cDNA or is used to identify desired sequences in genomic DNA libraries; RNA isolation; and The construction of cDNA and genomic libraries is well known to those of skill in the art. subel, or Sambrook, supra.)

[0073] Nucleic Acid Screening and Isolation Methods The sequences of polynucleotides used in the methods of the invention, such as those disclosed herein, Screening a cDNA or genomic library using sequence-based probes Probes can be used to hybridize to genomic DNA or cDNA sequences. By combining these sequences, it is possible to isolate homologous genes from the same or different organisms. Various degrees of hybridization stringency can be used in the Either the hybridization or the washing medium can be made stringent. It will be apparent that the more stringent the hybridization conditions, the greater the degree of duplex formation. There should be a high degree of complementarity between the probe and the target for formation to occur. The degree of confluency is affected by several variables, such as temperature, ionic strength, pH, and formamide. The hybridization rate can be controlled by one or more of the following: The stringency of the reaction can be determined, for example, by operating the formamide concentration in the range of 0% to 50%. This can be successfully altered by changing the polarity of the reaction solution through the action of The degree of complementarity (sequence identity) achieved may vary depending on the hybridization medium and / or washing medium. The degree of complementarity depends on the stringency of the nucleic acid. Optimally, the degree of complementarity is 100% or 70%. ~100%, or any range or value therein. However, Small differences in sequence among primers can affect the stability of hybridization and / or washing media. It should be understood that this can be compensated for by reducing the linkage.

[0074] Methods for amplifying RNA or DNA are well known in the art and are described herein. Based on the teachings and guidance provided, one can make use of the present invention without undue experimentation.

[0075] Known methods for amplifying DNA or RNA include the polymerase chain reaction (PCR). PCR) and related amplification processes (e.g., Mullis et al., U.S. Pat. Nos. 4,683,195, 4,683,202, and 4,683,203. ,800,159, 4,965,188, and Tabor et al., 4,795,69 Nos. 9 and 4,921,794, Innis' No. 5,142,033, Wils No. 5,122,464 to On et al., No. 5,091,310 to Innis, and No. 5,091,310 to Gyll. No. 5,066,584 to Ensten et al. and No. 4,889,810 to Gelfand et al. No. 8, Silver et al., No. 4,994,370, Biswas et al., No. 4,766,0 67, Ringold, 4,656,134), and double-stranded DNA. Use antisense RNA to a target sequence as a template for A synthesis A-mediated amplification (U.S. Pat. No. 5,130,238 to Malek et al., having the trade name NASBA) ), the entire contents of which are incorporated herein by reference. (See, e.g., Ausubel, supra, or Sambrook, supra.) Please refer to.)

[0076] For example, genomic DNA or cDNA may be amplified using polymerase chain reaction (PCR) techniques. Polynucleotides and related genes used in the methods of the invention can be isolated directly from the library. PCR and other in vitro amplification methods can also be used to amplify sequences, e.g., expression cloning a nucleic acid sequence encoding a protein of interest; detecting the desired m As probes for detecting the presence of RNA, for nucleic acid sequencing, or for other purposes It may be useful for producing nucleic acids for use as nucleic acids for in vitro amplification. Examples of techniques sufficient to guide the reader are found in Berger, supra; Sambrook, supra; and Ausubel and Mullis et al., U.S. Pat. No. 4,683,202 (1987) , and Innis, et al., PCR Protocols A Guide t ○Methods and Applications,Eds.,Academic Press Inc, San Diego, CA (1990). Genomic PCR Commercially available kits for amplification are known in the art. See GC Genomic PCR Kit (Clontech). In addition, For example, T4 gene 32 protein (Boehringer Mannheim) was used This can improve the yield of long PCR products.

[0077] Synthetic methods for constructing nucleic acids The isolated nucleic acids used in the methods of the present invention can also be prepared by direct chemical synthesis using known methods. They can also be prepared by chemical synthesis (see, for example, Ausubel et al., supra). Generally, by hybridization with a complementary sequence or by cleaving a single strand of a template A single strand that can be converted into double-stranded DNA by polymerization with a DNA polymerase used as a Those skilled in the art will appreciate that chemical synthesis of DNA has over 100 salts. Although the sequence of the base may be limited, longer sequences can be obtained by ligation of shorter sequences. You will realize that you can.

[0078] Recombinant Expression Cassettes The present invention uses recombinant expression cassettes that contain nucleic acids. At least one site has been identified using cDNA or genomic sequences encoding the antibody to be used. A recombinant expression cassette can be constructed that can be introduced into a desired host cell. An expression cassette typically directs transcription of a polynucleotide in an intended host cell. The term "heterologous" refers to a polynucleotide that is operably linked to a transcription initiation regulatory sequence that directs the transcription of the target gene. Both heterologous (i.e., endogenous) promoters can be used to direct expression of the nucleic acid. do.

[0079] In some embodiments, it functions as a promoter, enhancer, or other element. The isolated nucleic acid can be used to express a polypeptide of the invention to up- or down-regulate expression of the polynucleotide. By introducing a non-heterologous form of a nucleotide into the appropriate position (upstream, downstream, or within an intron), For example, endogenous genes can be modified by mutation, deletion and / or substitution in vivo or in vitro. The sex promoter can be varied.

[0080] Vectors and host cells The present invention relates to vectors comprising isolated nucleic acid molecules, recombinant vectors comprising isolated nucleic acid molecules, and expressing at least one antibody by recombinant techniques well known in the art. The production of IL-23 antibodies is also contemplated. See, e.g., Sambrook et al., supra; Ausu et al., supra. See bel et al., each of which is incorporated by reference in its entirety.

[0081] The polynucleotide may optionally be expressed as a vector containing a selectable marker for propagation in a host. Generally, plasmid vectors are prepared by calcium phosphate precipitation. The vector is introduced into a precipitate such as a vesicle or into a complex with a charged lipid. If the gene is a recombinant virus, it can be packaged in vitro using an appropriate packaging cell line and then transduced into the host cells. It can be introduced.

[0082] The DNA insert should be operably linked to a suitable promoter. Lactococci contain transcription initiation sites, transcription termination sites, and, within the transcribed region, ribosomal enzymes for translation. The coding portion of the mature transcript expressed by the construct preferably further comprises a In general, a start and stop codon (e.g., U In mammalian or eukaryotic cell expression, the translation begins with U AA and UAG are preferred.

[0083] The expression vector preferably contains at least one selectable marker, although this is optional. Such markers include, for example, methotrexate for eukaryotic cell culture. rexate, MTX), dihydrofolate reductase, DHFR, U.S. Patent Nos. 4,399,216, 4,634,665, and 4,656,134 No. 4,956,288, No. 5,149,636, No. 5,179,017 , ampicillin, neomycin (G418), mycophenolic acid or glutamine synthetase No. 5,122,464; U.S. Pat. No. 5,770,359; U.S. Pat. No. 5,770,359; U.S. Pat. No. 5, 827,739) Resistance genes and culture in E. coli and other bacteria or prokaryotes These include, but are not limited to, tetracycline or ampicillin resistance genes for (The above patents are incorporated herein by reference in their entirety.) Appropriate culture media and conditions are known in the art. Introduction of the vector construct into a host cell will be readily apparent to those skilled in the art. Calcium phosphate transfection, DEAE-dextran mediated transfection transfection, cationic lipid-mediated transfection, electroporation This can be achieved by injection, transduction, infection or other known methods. Sambrook, supra, chapters 1-4 and 16-18; Ausubel, supra, chapter 1 , Chapters 9, 13, 15, 16, and the like, in the art.

[0084] At least one antibody used in the methods of the invention may be a modified antibody, such as a fusion protein. The vector may be expressed in a form that includes not only a secretion signal 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 enhance its affinity for ribozyme during purification or subsequent processing. It can improve the stability and persistence in the host cell during processing and storage. Peptide moieties can also be added to the antibodies of the invention to facilitate purification. Such regions can be removed prior to final preparation of the fragment. The method is described in Sambrook, supra, Chapters 17.29-17.42 and 18.1-18. Many standard laboratory manuals, such as Chapter 74, Ausubel, supra, Chapters 16, 17 and 18, It's stated in the manual.

[0085] Those skilled in the art will appreciate that various methods are available to express nucleic acids encoding proteins for use in the methods of the present invention. Alternatively, the nucleic acid may be expressed intracellularly as a nucleic acid encoding an antibody. By switching on (by engineering) the gene in a host cell that contains the gene for the gene, the host cell Such methods can be expressed in cells. Nos. 5,641,670, 5,733,746, and 5,733,761. These methods are well known in the art, as described in the above publications, and are incorporated herein by reference in their entirety. will be incorporated into the subsection.

[0086] One example of a cell culture useful for producing an antibody, specified portion or variant thereof is a mammalian cell culture. Mammalian cell lines often take the form of a monolayer of cells, but suspensions of mammalian cells are Suspension or bioreactors can also be used. Intact glycosylated proteins can be expressed. Several suitable host cell lines have been developed in the art, including COS -1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-1 651), 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 Examples of such cells include Amer / 0-Ag14, 293 cells, and HeLa cells. ican Type Culture Collection,Manassas,Va (www.atcc.org). Preferred host cells include bone marrow Particularly preferred host cells include cells derived from the lymphatic system, such as lymphoma and lymphoma cells. P3X63Ag8.653 cells (ATCC registration no. CRL-1580) and SP2 / 0 -Ag14 cells (ATCC Accession No. CRL-1851). The recombinant cells are P3X63Ab8.653 or SP2 / 0-Ag14 cells.

[0087] Expression vectors for these cells include an origin of replication, a promoter (e.g., late or early SV 40 promoter, CMV promoter (U.S. Pat. No. 5,168,062, U.S. Pat. No. 5,3 85,839), HSV tk promoter, pgk (phosphoglycerate kinase) Promoter, EF-1α promoter (U.S. Patent No. 5,266,491), at least Another human immunoglobulin promoter, enhancer, and / or ribosome binding site positions, RNA splice sites, polyadenylation sites (e.g., SV40 large T Ag polyadenylation sites, addition sites), as well as processing information sites such as transcription termination sequences, but are not limited to these. The expression control sequences may include one or more of the expression control sequences described in, for example, Ausubel et al., supra. See Sambrook et al., supra. Useful for producing the nucleic acids or proteins of the invention Other such cells are known and / or may be used, for example, in the American Type The Culture Collection's catalog of cell lines and hybridomas (www atcc.org) or other known or commercial sources.

[0088] When eukaryotic host cells are used, polyadenylation or transcription termination sites are typically included in 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 be included as well. An example of a splicing sequence is the VP1 intron from SV40 (Sprague , et al., J. Virol. 45:773-781 (1983)). In addition, the As is known in the art, gene sequences for controlling replication within a host cell can be used. It can be incorporated into a vector.

[0089] Antibody purification Anti-IL-23 antibodies were purified using protein A, ammonium sulfate or ethanol precipitated, and extracted with acid. extraction, anion or cation exchange chromatography, phosphocellulose chromatography -, hydrophobic interaction chromatography, affinity chromatography, hydroxy These include luapatite chromatography and lectin chromatography. The polypeptides can be recovered and purified from recombinant cell culture by well-known methods, including but not limited to those described herein. High performance liquid chromatography (HPL C") can also be used for purification. See, e.g., Colligan, Current P rotocols in Immunology or Current Protocol s in Protein Science,John Wiley & Sons,N See, for example, chapters 1, 4, 6, 8, 9, and 10 of Y, NY (1997-2001). No. 6,399,623, each of which is incorporated herein by reference in its entirety.

[0090] The antibodies used in the method of the present invention may be naturally purified products, products produced by chemical synthesis techniques, or the like. and recombinantly produced from eukaryotic hosts, including, for example, yeast, higher plants, insects, and mammalian cells. Depending on the host utilized in the recombinant production procedure, The body may be glycosylated or non-glycosylated, but glycosylation is preferred. Such a method is described in Sambrook, supra, Sections 17.37-17.42. , Ausubel, supra, chapters 10, 12, 13, 16, 18, and 20, Col, supra Many standard studies, such as Ligan, Protein Science, Chapters 12-14, Laboratory manuals, all of which are incorporated herein by reference in their entireties.

[0091] Anti-IL-23 antibody The anti-IL-23 antibody of the present invention can be incorporated into an immunoglobulin molecule, At least a portion of, for example, but not limited to, at least one ligand binding portion nd binding portion, LBP), for example, but not limited to, heavy chain or light chain complementarity determination constant drive (CDR) or ligand-binding portion thereof, heavy or light chain variable region, framework region A region (e.g., FR1, FR2, FR3, FR4, or a fragment thereof, and optionally at least one substitution, insertion, or deletion), a heavy or light chain constant region (e.g., At least 1 C H1, Hinge 1, Hinge 2, Hinge 3, Hinge 4, C H 2 or C H 3 or a fragment thereof, and optionally containing at least one substitution, insertion, or deletion. ), or any portion thereof. may be human, mouse, rabbit, rat, rodent, primate, or any combination thereof. The subject matter may include or be derived from any mammal, including, but not limited to, a mammal such as a mammalian animal.

[0092] The isolated antibodies used in the methods of the present invention may be prepared by any suitable polynucleotide. The amino acid sequence of the antibody encoded and disclosed herein, or any isolated or prepared Preferably, the human antibody or antigen-binding fragment binds to human IL-23 and This partially or substantially neutralizes at least one biological activity of the protein. At least one biological activity of at least one IL-23 protein or fragment is retained. The antibodies, or specified portions or variants thereof, are preferably neutralizing, either partially or preferably substantially neutralizing. IL-23 is a cytoplasmic IL-23 molecule that binds to the IL-23 receptor and binds to the IL-23 protein or fragment, thereby mediating the 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 a neutralizing antibody that can be used to detect a specific antibody, depending on the assay. and about 20 to 120%, preferably at least about 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 9 Refers to antibodies that can inhibit IL-23-dependent activity by 7, 98, 99, 100% or more. The ability of an anti-IL-23 antibody to inhibit IL-23-dependent activity is preferably determined by the methods described herein. At least one suitable IL-23 as described and / or known in the art Human antibodies can be assessed by protein or receptor assays. IgA, IgM, IgE, IgD, etc.) or isotype, In one embodiment, the human antibody may comprise an IgG heavy chain or a defined fragment. fragments, such as IgG1, IgG2, IgG3 or IgG4 (e.g., γ1, γ2, γ3, γ4). Antibodies of this type are described herein. At least one human light chain, as described and / or known in the art (e.g., Transgenic mice or other human mice containing transgenes for IgG, IgA, and IgM. The present invention can be prepared by utilizing transgenic mammals other than the mouse. In one embodiment, the anti-IL-23 human antibody comprises an IgG1 heavy chain and an IgG1 light chain. .

[0093] The antibody is an antibody that binds at least one IL-23 protein, subunit, fragment, portion, or the like. It binds to at least one specified epitope specific for any combination of these. The at least one epitope comprises at least one polypeptide that comprises at least a portion of a protein. The epitope may comprise one antibody binding region, and the epitope is preferably located on the from at least one of an extracellular portion, a soluble portion, a hydrophilic portion, an outer portion, or a cytoplasmic portion; It is composed.

[0094] Generally, a human antibody or antigen-binding fragment contains at least one human complementarity determining region. (CDR1, CDR2, and CDR3) or at least one heavy chain variable region variant, and at least one human complementarity determining region (CDR1, CDR2, and CDR3) or at least The CDR sequences are derived from human germline variants of the light chain variable region. May be derived from a cell lineage sequence or may be an exact match to a germline sequence For example, CDRs derived from a synthetic library derived from original non-human CDRs are used. These CDRs can be modified by incorporating conservative substitutions from the original non-human sequence. In another particular embodiment, the antibody or antigen-binding portion or variant may be formed by at least one light chain CDR having a corresponding CDR1, 2 and / or 3 amino acid sequence; That is, an antigen-binding domain that contains at least a portion of CDR1, CDR2, and / or CDR3. It may have a merger region.

[0095] Such antibodies can be prepared using conventional techniques involving recombinant DNA technology to encode the antibodies (i.e. by preparing and expressing one or more) nucleic acid molecules, or by any other suitable method. By using the method, various portions of the antibody (e.g., CDRs, The polymeric framework can be prepared by chemically bonding together the polymerizable polymers.

[0096] The anti-IL-23 specific antibody has a heavy or light chain variable region having a defined amino acid sequence. For example, in a preferred embodiment, the antibody may include at least one of anti-IL-2 3. The antibody optionally comprises at least one heavy chain polypeptide having the amino acid sequence of SEQ ID NO: 106. Variable region, and / or optionally at least one having the amino acid sequence of SEQ ID NO: 116 The antibody binds to human IL-23 and has a defined heavy chain variable region. The antibodies comprising the light or light chain variable regions can be prepared using any of the methods known in the art and / or described herein. Phage display (Katsube, Y., et al., Int J Mol. Me d,1(5):863-868(1998)) or those who adopt transgenic animals. For example, functionally rearranged human IgG can be prepared using any suitable method, including the use of a method for the preparation of functionally rearranged human IgG. A human immunoglobulin heavy chain transgene capable of functionally rearrangement and a human immunoglobulin heavy chain transgene capable of functionally rearrangement. and a transgenic mouse comprising a transgene comprising DNA from the purine light chain locus. can be immunized with human IL-23 or a fragment thereof to induce the production of antibodies. In this case, antibody producing cells can be isolated and processed as described herein and / or As is known in the art, hybridomas or other immortalized antibody producing cells may be used. Alternatively, the antibody, specified portion or variant may be prepared in a suitable manner. In a suitable host cell, the encoding nucleic acid, or a portion thereof, can be used for expression.

[0097] The present invention also relates to amino acids in sequences which are substantially the same as the amino acid sequences described herein. The present invention also relates to antibodies, antigen-binding fragments, immunoglobulin chains and CDRs comprising the amino acid. Alternatively, such antibodies or antigen-binding fragments and antibodies comprising such chains or CDRs may be highly homologous. Harmonic (e.g., K D About 10 -9 M or less) and can bind to human IL-23. Amino acid sequences that are substantially the same as the sequences described herein include conservative amino acids. Conservative amino acid substitutions and sequences containing amino acid deletions and / or insertions are also included. The substitution is performed by substituting a nucleotide sequence that has chemical and / or physical properties (e.g., charge, structure, etc.) similar to those of the first amino acid. The substitution of a first amino acid with a second amino acid that has a specific structure, polarity, or hydrophobic / hydrophilicity. Conservative substitutions include, but are not limited to, replacing one amino acid with another amino acid within the following group: Substituting: lysine (K), arginine (R), and histidine (H); Paragine (D) and glutamate (E); asparagine (N), glutamine (Q) , serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; Aline (A), valine (V), leucine (L), isoleucine (I), proline (P), phenanthroline (P), nitrile (F), tryptophan (W), methionine (M), cysteine ​​(C), and and glycine (G); F, W, and Y; C, S, and T.

[0098] Amino acid code The amino acids constituting the anti-IL-23 antibodies of the present invention are often abbreviated. is its one-letter code, its three-letter code, its name, or the codon (compound) of three nucleotides. The amino acid sequence can be represented by a symbol (number or number) and is well known in the art. It is well understood (Alberts, B., et al., Molecular Biol ogy of The Cell,Third Ed.,Garland Publis (see, Fing, Inc., New York, 1994).

[0099] [Table 1] The anti-IL-23 antibodies used in the methods of the present invention are natural targeting antibodies, as specified herein. Substitution, deletion, or addition of one or more amino acids, either by natural mutation or by human manipulation. It may include addition.

[0100] The number of amino acid substitutions a skilled artisan would make depends on many factors, including those described above. In other words, the number of amino acid substitutions, insertions, or deletions in a given anti-IL-23 antibody, fragment, or variant. As specified herein, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, e.g., 1 to 3 Does not exceed 0 or any range or value therein.

[0101] Amino acids within the anti-IL-23 specific antibody that are functionally essential were identified by site-directed mutagenesis. or by methods known in the art, such as alanine scanning mutagenesis. can be identified (e.g., Ausubel, supra, Chapters 8, 15; Cunningham and Wells,Science 244:1081-10 85 (1989)). The latter procedure involves the mutation of each residue in the molecule with a single alanine. The resulting substitution mutant molecule is then, for example, but not limited to, The antibodies are tested for at least one biological activity, such as IL-23 neutralizing activity. The sites that are crucial for the synthesis of the nucleosomes have also been subjected to structural analysis, such as crystallization, nuclear magnetic resonance or photoaffinity labeling. (Smith, et al., J. Mol. Biol. 2002). 24:899-904 (1992) and de Vos, et al., Science 255:306-312(1992)).

[0102] The anti-IL-23 antibody may be selected from the group consisting of SEQ ID NOs: 5, 20, 44, 50, 56, and 73. At least one portion, sequence, selected from 5 to all of one adjacent amino acid The following may include, but are not limited to, a sequence, a column, or a combination.

[0103] The IL-23 antibody or specific portion or variant includes at least 3 of the above SEQ ID NO: 5 to 17 adjacent amino acids of the above SEQ ID NO: 10 adjacent amino acids, 5 to 11 adjacent amino acids of the above SEQ ID NO: 7 adjacent amino acids, at least one selected from 5 to 9 adjacent amino acids of the above sequence number It may include, but is not limited to, a single portion, sequence, or a combination.

[0104] The anti-IL-23 antibody further optionally comprises any one of the above SEQ ID NOs: 5, 17, 10, 11, 7, 9, At least one polypeptide having 70-100% of the adjacent amino acids of In one embodiment, the immunoglobulin chain, or a portion thereof (e.g., The amino acid sequence of the variable region (CDR) corresponds to at least one of the above SEQ ID NOs. The amino acid sequence of the chain has about 70 to 100% identity (e.g., 70, 71, 72, 73, 7 4, 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 For example, the light chain variable region has the amino acid sequence can be compared to the above SEQ ID NOs, or the amino acid sequence of the heavy chain CDR3 can be compared to the above SEQ ID NOs. Preferably, the amino acid sequence is 70 to 100% identical (i.e. , 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or any of the above Any range or value of the above may be calculated using a suitable computer, as known in the art. The time is determined using a data algorithm.

[0105] As known in the art, "identity" is determined by comparing sequences. The relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by the In the art, "identity" is also determined by the match between such linear sequences. "Sequence relatedness" refers to the degree of sequence relatedness between polypeptide or polynucleotide sequences, as determined by the "Identity" and "similarity" are defined as iology,Lesk,AM,ed.,Oxford University P ress, New York, 1988, Biocomputing:Informat. ics and Genome Projects, Smith, DW, ed., A Academic Press, New York, 1993, Computer Ana Analysis of Sequence Data,Part I,Griffin,A. M.,and Griffin,HG,eds.,Humana Press,Ne. w Jersey,1994, Sequence Analysis in Molec ular Biology, von Heinje, G., Academic Pres. s, 1987, and Sequence Analysis Primer, Gribsk ov, M. and Devereux, J., eds., M Stockton Pre. ss, New York, 1991, and Carillo, H., and Lipma n, D., Siam J. Applied Math., 48:1073 (1988). The methods can be readily performed by known methods, including but not limited to those described herein. In addition, the percent identity value can be calculated using the Vector NTI S A component of Suite 8.0 (Informax, Frederick, MD) Amino acids and nucleotides generated using default settings for lignX. This can be obtained from sequence alignment.

[0106] Preferred methods to determine identity are those that give the largest match between the sequences tested. Methods to determine identity and similarity are designed using publicly available computer programs. The method for determining identity and similarity between two sequences is codified in the NIH Standards Track [Page 11] RFC 2411 Sequence Identification and Similarity Index (SIGMA). A preferred computer program method for doing this is the GCG program package (Devere ux, J., et al., Nucleic Acids Research 12(1 ):387(1984)), BLASTP, BLASTN, and FASTA (Atsch ul,SFet al.,J.Molec.Biol.215:403-410(1 990)). The BLAST X program is available from NCBI and and other sources (BLAST Manual, Altschul, S., et al., N CBINLM NIH Bethesda,Md.20894:Altschul,S. , et al., J. Mol. Biol. 215:403-410 (1990) The well-known Smith Waterman algorithm can also be used to determine identity. It can be used to

[0107] Preferred parameters for polypeptide sequence comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Bi ol.48:443-453(1970)Comparison matrix:BLOSSUM62 fr om Hentikoff and Hentikoff,Proc.Natl.Aca d.Sci,USA.89:10915-10919(1992), Gap penalty: 12 Gap length penalty: 4 A useful program for these parameters is Genetics Computer G Publicly available as the "Gap" program from roup, Madison Wis. The above parameters are the default parameters for peptide sequence comparison (terminal (Just as there is no penalty for caps.)

[0108] Preferred parameters for polynucleotide comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Bi ol.48:443-453(1970) Comparison matrix: match = +10, mismatch = 0 Gap penalty: 50 Gap length penalty: 3 " from Genetics Computer Group, Madison Wis. These are available as the "Gap" program, which uses the default parameters for nucleic acid sequence comparison. It is.

[0109] By way of example, a polynucleotide sequence can be identical to another sequence, i.e., 100% identical. may contain one or up to a certain integer number of nucleotide changes compared to the reference sequence. Such alterations include at least one nucleotide deletion, substitution (including transitions and transversions), or an insertion, wherein the alteration is at the 5' or 3' end of the reference nucleotide sequence. It may occur at the end positions or anywhere between these end positions and may be between nucleotides of the reference sequence. Nucleotides may be present individually or interspersed in one or more adjacent groups within the reference sequence. The number of nucleotide changes corresponds to the total number of nucleotides in the sequence and the corresponding percent identity number. Multiply by the percentage (divided by 100) and subtract that product from the total number of nucleotides in the sequence. Pulling or By n.sub.n.ltorsim.x.sub.n-(x.sub.ny) It was decided, where n.sub.n is the number of nucleotide changes and x.sub.n is the number of nucleotide changes in the sequence. The total number of leucocytes, y is, for example, 0.70 for 70% and 0 for 80%. .80, 0.85 for 85%, 0.90 for 90%, and 0. 95, and any non-integer product of x.sub.n and y is x.sub.n to y. Round down to the nearest integer before subtracting.

[0110] Alterations in the polynucleotide sequence encoding the above SEQ ID NOs may be made without restriction to the coding sequence. create a sense, missense, or frameshift mutation, thereby The polypeptide encoded by the polynucleotide can then be modified. Similarly, the polypeptide sequence may be identical to the reference sequence of the above SEQ ID NO: 10 A specific sequence is compared to the reference sequence such that it is 0% identical or the identity is less than 100%. Such modifications may include the deletion of at least one amino acid, up to a certain integer. The alteration is selected from the group consisting of a deletion, a substitution (including conservative and non-conservative substitutions), or an insertion. , at the amino or carboxy terminal positions of the reference polypeptide sequence, or at these terminal positions and may occur anywhere between the amino acids of the reference sequence individually or at one or more positions within the reference sequence. The amino acid changes for a given % identity may be interspersed in any of the flanking groups. The numbers represent the total number of amino acids in the sequence shown, and the corresponding percent identity. (divided by 100) and subtract the product from the total number of amino acids in the sequence. Or, By n.sub.a.ltorsim.x.sub.a-(x.sub.ay) It was decided, where n.sub.a is the number of amino acid changes and x.sub.a is the is the total number of amino acids, and y is, for example, 0.70 for 70% and 0 for 80%. .80 for 85%, 0.85 for 85%, etc. for any integers x.sub.a and y. Any missing products are truncated to the nearest integer before being subtracted from x.sub.a.

[0111] Exemplary heavy and light chain variable region sequences, and portions thereof, are set forth in the SEQ ID NOs. The antibodies of the invention or specified variants thereof may be any number of adjacent amino acids from an antibody of the invention. The number of adjacent acid residues may be 10-100% of the number of adjacent residues in the anti-IL-23 antibody. Optionally, the subsequence of contiguous amino acids is selected from the group of integers consisting of at least Also 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 therein. Further, the number of such subsequences is at least 1 to 2, such as 2, 3, 4, or 5. It may be any integer selected from the group consisting of 0.

[0112] As will be apparent to one of skill in the art, the present invention includes at least one biologically active compound of the present invention. Biologically active antibodies include antibodies that are naturally occurring (non-synthetic), endogenous, or related, and At least 20%, 30%, or 40% of that of the known antibody, and preferably at least At least 50%, 60%, or 70%, and most preferably at least 80%, 90%, or 9 having a specific activity of 5% to 100% or more (including but not limited to up to 10 times the specific activity). Methods for assaying and quantitatively measuring enzyme activity and substrate specificity are well known to those of skill in the art.

[0113] In another aspect, the present invention provides a method for the preparation of a compound according to any one of the preceding claims, wherein the compound is modified by the covalent attachment of an organic moiety. The present invention relates to human antibodies and antigen-binding fragments that have been modified to have improved pharmacokinetic properties. (e.g., increased serum half-life in vivo). The organic moiety can be a linear or branched hydrophilic polymer group, a fatty acid group, or a fatty acid ester. In certain embodiments, the hydrophilic polymeric group can be a tertiary ether group. 0 to about 120,000 daltons, and polyalkane glycols (e.g., polyethylene glycols). Polyethylene glycol (PEG), polypropylene glycol polyvinylpyrrolidone (PPG), carbohydrate polymers, amino acid polymers or polyvinylpyrrolidone (PPG) and the fatty acid group or fatty acid ester group contains from about 8 to about 40 carbon atoms. obtain.

[0114] The modified antibodies and antigen-binding fragments are directly or indirectly covalently attached to the antibody. The antibody or antigen-binding fragment of the present invention may contain one or more organic moieties that are attached to the antibody or antigen-binding fragment of the present invention. Each organic moiety is independently a hydrophilic polymeric group, a fatty acid group, or a fatty acid ester group. As used herein, the term "fatty acid" refers to mono- and di-carboxylic acids. As used herein, the term "hydrophilic polymeric group" includes octane, It refers to an organic polymer that is more soluble in water than in water. For example, polylysine is Therefore, antibodies modified with covalent polylysine have a higher solubility in water than Suitable hydrophilic polymers for modifying the antibodies of the invention include linear or branched hydrophilic polymers. They may be branched, for example polyalkane glycols (e.g., PEG, monomethoxy-poly Ethylene glycol (monomethoxy-polyethylene glycol, mPEG, PPG, etc.), Carbohydrates (e.g., dextran, cellulose, oligosaccharides, polysaccharides, etc.), hydrophilic amino acids Polymers (e.g., polylysine, polyarginine, polyaspartic acid, etc.), poly Glucan oxides (e.g., polyethylene oxide, polypropylene oxide, etc.), and Preferably, the hydrophilic polymer modifying the antibody of the present invention comprises As an individual molecular entity, it has a molecular weight of about 800 to about 150,000 daltons. For example, PEG 5000 and PEG 20,000 can be used, and the subscripts indicate the polymer The average molecular weight (Daltons) of the hydrophilic polymer group is 1 to about 6 alkyl groups, fatty acids, It can be substituted with an acid group or a fatty acid ester group. The hydrophilic polymers can be prepared by utilizing any suitable method. For example, a polymer containing amine groups can be linked to the carboxylate of a fatty acid or fatty acid ester. and activating carboxylates (e.g., N,N- (activated with carbonyldiimidazole) is linked to the hydroxyl groups on the polymer. It is possible to do so.

[0115] The fatty acids and fatty acid esters suitable for modifying the antibodies of the invention may be saturated. Suitable lipids for modifying the antibodies of the invention may contain one or more unsaturated units. Examples of fatty acids include n-dodecanoic acid salts (C 12 , laurate), n-tetradecane Acid acid (C 14 , myristate), n-octadecanoate (C 18 , stearates), n-Eicosanoic acid salt (C 20 , arachidate), n-docosanoate (C 22 , behenic acid ), n-triacontanoate (C 30 ), n-tetracontanoate (C 40 ), cis-Δ 9-Octadecanoic acid salt (C 18 oleate), all cis-Δ5,8,11,14- Eicosatetraenoic acid salt (C 20 , arachidonate), octanedioic acid, tetradecane Suitable fatty acid esters include ethyl ester, octadecanedioic acid, docosanedioic acid, and the like. The esters include monoesters of dicarboxylic acids containing straight or branched chain lower alkyl groups. The lower alkyl group may contain from 1 to about 12, preferably from 1 to about 6, carbon atoms.

[0116] The modified human antibodies and antigen-binding fragments can be subjected to any suitable treatment, such as by reacting them with one or more modifying agents. As used herein, the term "modifying agent" means , a suitable organic group (e.g., a hydrophilic polymer, a fatty acid, a fatty acid ester) containing an activating group. An "activating group" means a group that, under appropriate conditions, reacts with a second chemical group, thereby forming a modifying agent. A chemical moiety or functional group capable of forming a covalent bond between a first chemical group and a second chemical group. For example, amine-reactive activating groups include tosylate, mesylate, halo (chloro, bromo, Electrophilic groups such as fluoro and iodo, N-hydroxysuccinimidyl esters (N-hydrox Activated groups that can react with thiols and Examples of suitable aryl groups include maleimide, iodoacetyl, acryloyl, and pyridyl disulfides. 5-thiol-2-nitrobenzoic acid thiol (TNB-thiol) and the like. The aldehyde functional group can be linked to an amine- or hydrazide-containing molecule, and The dido group reacts with a trivalent phosphorus group to form a phosphoramidate or phosphorimide bond. Suitable methods for introducing activating groups into molecules are known in the art. It is known (e.g., Hermanson, GT, Bioconjugate Te chniques, Academic Press: San Diego, CA (199 6) The activating group can be an organic group (e.g., a hydrophilic polymer, a fatty acid, ester) directly or via a linker moiety, e.g., a divalent C-C 12 Group (where 1 one or more carbon atoms may be replaced by heteroatoms such as oxygen, nitrogen, or sulfur, Suitable linker moieties include, for example, tetraethylene glycol, - (CH2)3-, -NH-(CH2)6-NH-, -(CH2)2-NH- and -CH2 The linker portion includes -O-CH2-CH2-O-CH2-CH2-O-CH-NH-. The modifying agent may be, for example, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. In the presence of (ethyl-3-(3-dimethylaminopropyl) carbodiimide, EDC), mono-Boc -Alkyl diamines (e.g., mono-Boc-ethylenediamine, mono-Boc-diamine By reacting hexane with fatty acids, free amines and fatty acid carboxylates are produced. The Boc protecting group can be converted to trifluoroacetate by forming an amide bond between The product was removed by treatment with trifluoroacetic acid (TFA) as described or exposing a primary amine that can be coupled to another carboxylate, such as This is reacted with maleic anhydride and the resulting product is cyclized to give the activated maleimide fatty acid. Derivatives can be produced using the methods described in, for example, US Pat. (See Thompson et al., International Publication WO 92 / 16221, which is incorporated herein by reference).

[0117] The modified antibodies can be prepared by reacting a human antibody or antigen-binding fragment with a modifying agent. For example, the organic moiety can be produced by an amine-reactive modifying agent, e.g., PEG. NHS esters of the compounds can be used to conjugate to antibodies in a non-site-specific manner. Disulfide bonds (e.g., intrachain disulfide bonds) of antibodies or antigen-binding fragments can be (a) preparing a modified human antibody or antigen-binding fragment thereof; In this case, the reduced antibody or antigen-binding fragment can be subjected to thiol-reactive modification. The antibody of the present invention can be reacted with a decorating agent to produce a modified antibody of the present invention. Modified human antibodies and antigen-binding fragments containing organic moieties attached to specific sites of the antibody are known as reverse transcription antibodies. Protein degradation (Fisch et al., Bioconjugate Chem., 3 :147-153(1992), Werlen et al., Bioconjugat e Chem.,5:411-417(1994), Kumaran et al.,P rotein Sci.6(10):2233-2241(1997), Itoh et al. al.,Bioorg.Chem.,24(1):59-68(1996), Cape llas et al.,Biotechnol.Bioeng.,56(4):456 -463 (1997)) and Hermanson, GT, Bioconjugate Techniques, Academic Press: San Diego, CA( The compounds can be prepared using any suitable method, such as that described in (1996).

[0118] The methods of the present invention also include those described herein and / or known in the art. At least one of the following is provided in a composition, mixture, or form that is not naturally occurring: one, at least two, at least three, at least four, at least five, at least six Also used are anti-IL-23 antibody compositions comprising one or more anti-IL-23 antibodies. The amino acid sequence is 70-100% of the adjacent amino acids of the above sequence, or a specified fragment, domain, or or a variant thereof. One or two full-length, C- and / or N-terminal deletion variants, domains, fragments, or specific Preferred anti-IL23 antibody compositions include, for example, non-naturally occurring compositions that include the modified variants. , an anti-IL23 antibody sequence described herein which is 70 to 100% of the above-mentioned SEQ ID NO, or At least one CDR or LBP-containing fragment, domain, or variant of The desired portion includes at least one or two full-length, fragments, domains, or variants. More preferred compositions include those that contain, for example, 70 to 100% of the above SEQ ID NOs., or a combination thereof. Contains 40-99% of at least one of the fragments, domains, or variants The percentages of such compositions may be as known in the art or as described herein. As described above, the weight, volume, concentration, molarity, or liquid or anhydrous solutions may be used. concentration as a mixture, suspension, emulsion, particle, powder, or colloid This is due to.

[0119] Antibody compositions containing further therapeutically active ingredients The antibody compositions used in the methods of the present invention may optionally further comprise an anti-infective agent, a cardiovascular scular (CV) acting drugs, central nervous system (CNS) drugs, autonomic nervous autonomic nervous system (ANS) drugs, respiratory drugs, gastrointestinal, GI ) Vascular activating agents, hormonal agents, fluid or electrolyte balancing agents, hemoactive agents, antitumor agents, immunomodulatory agents, A small amount of medicine selected from at least one of the following: eye, ear or nose medicines, topical medicines, nutritional medicines, etc. Such agents may include an effective amount of at least one compound or protein. The respective formulations, indications, dosages, and administrations set forth herein are well known in the art. (See, for example, Nursing 2001 Handbook of Drugs, 2 1 stedition,Springhouse Corp.,Springhouse e, PA, 2001, Health Professional's Drug Gui. de 2001, ed., Shannon, Wilson, Stang, Prentic. e-Hall, Inc., Upper Saddle River, NJ, Pharmco therapy Handbook,Wells et al.,Appleton&L see, e.g., Ange, Stamford, CT, each of which is incorporated herein by reference. (included).

[0120] Examples of drugs that can be combined with the antibodies of the methods of the invention include anti-infective drugs, such as ampicillin, or at least one antiprotozoal, anthelmintic, antifungal, antimalarial, antituberculous, or At least one antimycobacterial drug, aminoglycoside, penicillin, cephalosporin, tetracycline Lacyclines, sulfonamides, fluoroquinolones, antivirals, macrolides anti-infectives The hormonal drug may be at least one selected from the group consisting of steroids, anti-inflammatory drugs, and various anti-infective drugs. steroids, androgens, or at least one anabolic steroid, estrogens, or at least one progestin, gonadotropin, antidiabetic agent, or A type of glucagon, thyroid hormone, thyroid hormone antagonist, pituitary hormone, and At least one selected from the group consisting of parathyroid mimetics and at least one cephalosporin. Phosphorus is cefaclor, cefadroxil, cefazolin sodium, cefdinir, hydrochloride Cefepime, cefixime, cefmetazole sodium, cefonicid sodium, cef Operazon sodium, cefotaxime sodium, cefotetan disodium, cefo Cefpodoxime sodium, cefpodoxime proxetil, cefprozil, ceftazidime, cef Thibuten, ceftizoxime sodium, ceftriaxone sodium, cefuroxime aki Cetyl, Cefuroxime Sodium, Cephalexin Hydrochloride, Cephalexin Monohydrate, Cef The active ingredient may be at least one selected from the group consisting of azithromycin and loracarbef.

[0121] At least one corticosteroid is betamethasone, betamethasone acetate, Methasone or betamethasone sodium phosphate, betamethasone sodium phosphate, cholesteryl acetate Tisone, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, acetate Fludrocortisone, Hydrocortisone, Hydrocortisone Acetate, Hydrocortisone Cypionate Hydrocortisone, Hydrocortisone Sodium Phosphate, Hydrocortisone Sodium Succinate, Methionine Tilprednisolone, Methylprednisolone Acetate, Methylprednisolone Sodium Succinate um, prednisolone, prednisolone acetate, prednisolone sodium phosphate, tebutol Prednisolone, prednisone, triamcinolone, triamcinolone acetonide, and and triamcinolone diacetate. Androgenic or anabolic steroids include danazol, fluoxymesterone, Methyltestosterone, Nandrolone Decanoate, Nandrolone Phenpropionate, Tes Testosterone, Testosterone Cypionate, Testosterone Enanthate, Testosterone Propionate The testosterone may be at least one selected from the group consisting of testosterone transdermal agents.

[0122] At least one immunosuppressant is selected from the group consisting of azathioprine, basiliximab, cyclosporine, Daclizumab, lymphocyte immunoglobulin, muromonab-CD3, mycophenolate mofetil mycophenolate mofetil, mycophenolate mofetil hydrochloride, sirolimus, and tacrolimus. There can be at least one type.

[0123] At least one topical anti-infective agent is selected from the group consisting of acyclovir, amphotericin B, azelaic acid cyclovir, and the like. Ream, bacitracin, butoconazole nitrate, clindamycin phosphate, clotrimazole nitrate, econazole, erythromycin, gentamicin sulfate, ketoconazole, acetate Mafenide, metronidazole (topical), miconazole nitrate, mupirocin, naphthyl hydrochloride , neomycin sulfate, nitrofurazone, nystatin, silver sulfadiazine, tetrahydrofuran Rubinafine, terconazole, tetracycline hydrochloride, tioconazole, and tolnaf At least one scabicide or at least one tate. The pediculicide is selected from crotamiton, lindane, permethrin, and pyrethrins. The at least one topical corticosteroid may be dipropionate. Betamethasone valerate, clobetasol propionate, desonide, desoxyglucose Ximethasone, dexamethasone, dexamethasone sodium phosphate, diflorasone diacetate , fluocinolone acetonide, fluocinonide, flurandrenolide, flupropionate Ticasonide, halcionide, hydrocortisone, hydrocortisone acetate, butyrate Hydrocortisone valerate, hydrocortisone valerate, mometasone furoate, and triamcinol The at least one selected from the group consisting of acetonides and acetonides (see, for example, Nursing 2 See pages 1098-1136 of the 001 Drug Handbook.)

[0124] The anti-IL-23 antibody composition is intended to be used to administer to cells, tissues, organs, or tissues in need of such modulation, treatment, or therapy. At least one anti-IL-23 antibody is contacted or administered to a physician, animal, or patient. and optionally further comprising at least one TNF antagonist (e.g., but not limited to, T NF chemical or protein antagonists, TNF monoclonal or polyclonal antibodies or fragments thereof, soluble TNF receptors (e.g., p55, p70, or p85) or fragments thereof, Fusion polypeptides, or small molecule TNF antagonists, such as TNF binding protein I or II (TBP-1 or TBP-II), nerelimonmab, infliximab Mab, etanercept, CDP-571, CDP-870, Afelimoma antirheumatic drugs (e.g. methotrexate, auranofin, Aurothioglucose, azathioprine, etanercept, sodium gold thiomalate, Hydroxychloroquine sulfate, leflunomide, sulfasalazine), immunomodulators, immunoglobulins robrin, immunosuppressants (e.g., basiliximab, cyclosporine, daclizumab), Any suitable agent including at least one selected from the group consisting of cytokines and cytokine antagonists. The composition may further comprise at least one of a composition or a pharmaceutical composition in an effective amount. Non-limiting examples of cytokines include IL-1 to IL-23 (e.g., IL-1 The appropriate dose may include, but is not limited to, any of the following: It is well known in the art. For example, see Wells et al., eds., Pha rmacotherapy handbook, 2 nd Edition, Applet on and Lange, Stamford, CT (2000), PDR Pharm. acopoeia, Tarascon Pocket Pharmacopoeia 2 000,Deluxe Edition,Tarascon Publishing,L See, for example, Oma Linda, CA (2000), each of which is incorporated by reference in its entirety. The body is incorporated herein.

[0125] The anti-IL-23 antibody compound, composition, or mixture used in the methods of the present invention may further comprise dilutions. agents, binders, stabilizers, buffers, salts, lipophilic solvents, preservatives, adjuvants, etc. The composition may include at least one of any suitable auxiliary agent, including but not limited to, pharmaceutically acceptable Non-limiting examples of and methods for preparing such sterile solutions are described in the art. The method is well known in the art and is described, for example, in Gennaro, Ed., Remington's Pharmaceutical Sciences, 18 th Edition,Ma ck Publishing Co. (Easton, PA), 1990, etc. However, the present invention is not limited to the above. The method of administration, solubility, and / or stability of the anti-IL-23 antibody, fragment, or variant composition. A suitable pharma- ceutically acceptable carrier can be routinely selected.

[0126] Pharmaceutical excipients and additives useful in the present compositions include, but are not limited to, protein Proteins, peptides, amino acids, lipids and carbohydrates (e.g., monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and Sugars including oligosaccharides, alditols, aldonic acids, esterified sugars, and other derivative sugars, and polysaccharides or sugar polymers), which may be present alone or in combination. , alone or in combination, at 1 to 99.99% by weight or volume. The excipients include serum albumin such as human serum albumin (HSA). Recombinant human albumin (rHA), gelatin, Representative amino acids / antibody components that can also function in buffering capacity include Alanine, glycine, arginine, betaine, histidine, glutamic acid, asparagus Glycine, Cysteine, Lysine, Leucine, Isoleucine, Valine, Methionine, Phenyl Alanine, aspartame, etc. One of the preferred amino acids is glycine. .

[0127] Carbohydrate excipients suitable for use in the present invention include, for example, fructose, maltose, Monosaccharides such as saccharides like sucrose, galactose, glucose, D-mannose, sorbose, lactose , disaccharides such as sucrose, trehalose, cellobiose, raffinose, melezitose , maltodextrin, dextran, starches and other polysaccharides, mannitol, xylitol Tol, maltitol, lactitol, xylitol, sorbitol (glucitol), mi Preferred carbohydrates for use in the present invention include alditols such as inositol. The excipients are mannitol, trehalose, and raffinose.

[0128] The anti-IL-23 antibody composition may also include a buffer or pH adjusting agent, typically The buffer is a salt prepared from an organic acid or base. Representative buffers include citric acid, Acids, such as salts of ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid. The composition may be any organic acid salt, Tris, tromethamine hydrochloride, or phosphate buffer. Preferred buffers for use in are organic acid salts such as citric acid.

[0129] In addition, the anti-IL-23 antibody composition may be prepared by mixing 100% polyvinylpyrrolidone, Ficoll (a polymeric sugar) and 100% polyvinylpyrrolidone. ), dextrates (e.g., 2-hydroxypropyl-β-cyclodextrin, etc. Cyclodextrin), polyethylene glycol, flavoring agent, antibacterial agent, sweetener, antioxidant, Antistatic agents, surfactants (e.g., polyethersulfones such as "TWEEN 20" and "TWEEN 80") resorbates), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol may include polymeric excipients / additives such as chelating agents (e.g., EDTA) .

[0130] These and other compounds suitable for use in anti-IL-23 antibody, portion or variant compositions according to the invention. and additional known pharmaceutical excipients and / or additives are known in the art, e.g. For example, "Remington: The Science & Practice of Ph "armacy", 19 th ed., 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. The carrier or excipient materials include carbohydrates (e.g., monosaccharides and alditols) and buffers (e.g., chlo- charides ). An exemplary carrier molecule is the mucopolysaccharide, hyaluronic acid, which These may be useful for intra-articular delivery.

[0131] formulation As mentioned above, the present invention preferably comprises a phosphate salt solution containing saline or a selected salt. Buffered, stable formulations, as well as storage solutions and formulations containing preservatives, and pharma- ceutical Preferred for pharmaceutical or veterinary use are those comprising at least one anti-IL23 antibody in an acceptable formulation. The present invention provides a versatile and versatile preservative formulation comprising at least one known preservative in an aqueous diluent. , i.e., at least one of phenol, m-cresol, p-cresol, o-cresol Benzyl alcohol, phenylmercuric nitrate, phenoxyethanol Alcohol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate), Alkylparaben (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, salt benzethonium chloride, sodium dehydroacetate, and thimerosal, or a mixture thereof The composition further comprises a preservative, optionally selected from the group consisting of: As shown in the table, 0.001 to 5%, or 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, 4.0, 4.3, 4.5, 4.6, 4.7 , 4.8, 4.9, or any range or value therein. However, any suitable concentration or mixture of any range or value therein may be used. Non-exhaustive 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 to 3% benzyl alcohol (e.g. For example, 0.5, 0.9, 1.1, 1.5, 1.9, 2.0, 2.5%), 0.001 to 0. 5% thimerosal (e.g., 0.005, 0.01), 0.001-2.0% pheno rule (e.g., 0.05, 0.25, 0.28, 0.5, 0.9, 1.0%), 0.00 0.5-1.0% alkylparaben(s) (e.g., 0.00075, 0.0009 , 0.001, 0.002, 0.005, 0.0075, 0.009, 0.01, 0.0 2, 0.05, 0.075, 0.09, 0.1, 0.2, 0.3, 0.5, 0.75, 0 .9, 1.0%) etc.

[0132] As mentioned above, the method of the present invention comprises the steps of: (a) providing a packaging material and, optionally, a buffer formulated in an aqueous diluent; A small amount of solution containing at least one anti-IL-23 specific antibody with a buffer and / or preservative. The packaging material contains at least one vial of the solution, and the solution is stored in a container for one, two, three or four days. , 4, 5, 6, 9, 12, 18, 20, 24, 30, 36, 40, 48, 54, 60, 6 6. The label includes a description that the composition can be preserved for 72 hours or more. A packaging material, a first vial containing a lyophilized anti-IL-23 specific antibody, and a formulated and a second vial containing an aqueous diluent of the buffer or preservative, The packaging material is designed to preserve the anti-IL-23 specific antibody reconstituted in an aqueous diluent for more than 24 hours. The composition includes a label instructing the patient to form a sustained solution.

[0133] The anti-IL-23 specific antibodies used in accordance with the present invention may be any of those described herein or of the It is known in the art that the polypeptides can be produced from mammalian cells or transgenic preparations. The polypeptides may be produced by recombinant means, including recombinant human polypeptides, or may be purified from other biological sources.

[0134] The range of anti-IL-23 specific antibodies is approximately 1.0 μg / mL when reconstituted in the wet / dry system. It is contained in an amount that gives a concentration of about 1000 mg / mL to about 1000 mg / mL, but lower and higher concentrations are also available. The drug may be administered at any temperature, depending on the intended delivery vehicle. For example, in a solution formulation, the drug may be administered by transdermal penetration. This is distinct from pulmonary, transmucosal, or osmotic or micropump methods.

[0135] Preferably, the aqueous diluent optionally further comprises a pharma- ceutically acceptable preservative. New preservatives include phenol, m-cresol, p-cresol, o-cresol, and chloroform. Croxysol, benzyl alcohol, alkylparaben (methyl, ethyl, propyl, butyl ethyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate and Thimerosal or mixtures thereof. The concentration of preservative used is sufficient to produce an antimicrobial effect. The amount of preservative used will vary depending on the preservative used and can be readily determined by one of skill in the art.

[0136] Other excipients, such as isotonicity agents, buffers, antioxidants, and preservative enhancers, are optional. Optionally and preferably, an isotonic agent such as glycerin may be added to the diluent. Preferably, a physiologically tolerable buffer is added to improve the stability. The formulations provide a pH control ranging from about pH 4 to about pH 10, and preferably from about pH 5 to about pH A wide range of pH values, such as the pH range of H9, and most preferably the pH range of about 6.0 to about 8.0, Preferably, the formulation of the present invention has a pH of about 6.8 to about 7.8. Suitable buffers include phosphate buffers, most preferably sodium phosphate, especially sodium phosphate. Contains phosphate buffered saline (PBS).

[0137] If desired, other additives, such as Tween 20 (polyoxyethylene (20) sorbitol), may be added. monolaurate), Tween 40 (Polyoxyethylene (20) Sorbitan Monopal mitate), Tween 80 (Polyoxyethylene (20) sorbitan monooleate) , Pluronic F68 (polyoxyethylene polyoxypropylene block copolymer) Pharmacologically acceptable solubilizers, such as PEG (polyethylene glycol), and PEG (polyethylene glycol). , or polysorbate 20 or 80 or poloxamer 184 or 188, Plu Non-ionic surfactants such as bronic® polyols, other The formulation or composition may include a block copolymer and a chelating agent such as EDTA and EGTA. Addition of these additives can reduce aggregation. This is particularly useful when a pump or plastic container is used for the purpose. The presence of a surfactant reduces the tendency of proteins to aggregate.

[0138] The formulation comprises at least one anti-IL-23 specific antibody and phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkylpa Raben (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzyl chloride from the group consisting of thimerosal, sodium dehydroacetate, and thimerosal or mixtures thereof and a preservative selected from the group consisting of: The present invention relates to a method for the preparation of a pharmaceutical composition comprising administering to a patient a pharmaceutical composition comprising at least one anti-IL-23 specific antibody and a preservative in an aqueous diluent. Mixing is carried out 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 added to a desired concentration. and a desired preservative in a sufficient amount of buffer solution to provide the protein and preservative. Variations on this process will be recognized by those skilled in the art. For example, The order of addition of the ingredients, whether or not additional excipients are used, and the temperature and pH at which the formulation is prepared can all be determined by the These are factors that can be optimized with respect to the dosage concentration and means of administration.

[0139] The formulation may be prepared as a clear solution or in the form of a mixture of water, preservatives and / or excipients, preferably phosphate. Salt buffer and / or saline, and a second solution containing the selected salt in an aqueous diluent. A combination comprising a vial of lyophilized anti-IL-23 specific antibody, reconstituted in a vial of It can be provided to the patient as a single solution vial or as a dual vial. Any dual vial requiring configuration can be reused multiple times and can be used in single or multiple This could satisfy many patient treatment cycles and therefore provide a more convenient treatment option than currently available. A therapeutic regimen can be provided.

[0140] The product is useful for administration over periods ranging from immediate to 24 hours or more. Thus, the products claimed in accordance with the present invention provide significant benefits to patients. Optionally, safe storage at temperatures between about 2°C and about 40°C will maintain the biological activity of the protein for a long period of time. The packaging label therefore indicates that the solution can be stored for 6, 12, 18, 24, 36, Demonstrate that the product can be retained and / or used for periods of 48, 72, or 96 hours or more. When using stored diluents, such labels shall include a maximum of 1 to 1 This may include use for up to 2 months, 6 months, 1.5 years, and / or 2 years.

[0141] Solutions of anti-IL-23 specific antibodies are prepared by mixing at least one antibody in an aqueous diluent. The mixture can be prepared by a process comprising: To prepare a suitable diluent, for example, a small amount of water or buffer may be added. At least one antibody is added to the culture medium at a desired concentration of protein, and optionally a preservative or buffer. Variations of this process will be recognized by those of skill in the art. For example, the order of addition of the components, whether additional excipients are used, and the time and place of preparation of the formulation. The temperature and pH of the above can all be optimized for the dosage concentration and means of administration used. It is a factor.

[0142] The claimed product is available as a clear solution or in a second vial containing an aqueous diluent. The method includes the steps of: (a) administering to a patient a vial of at least one lyophilized anti-IL-23 specific antibody that is reconstituted in a lyophilized solution; It can be provided to patients as a single solution vial or as a combination vial that requires reconstitution. Each required dual vial can be reused multiple times and can be used for single or multiple patients. A treatment cycle can be filled, thus providing a more convenient treatment regimen than is currently available. Provides a.

[0143] The claimed product is a clear solution or a second vial containing an aqueous diluent. a combination comprising at least one lyophilized vial of an anti-IL-23 specific antibody, By providing the vials to pharmacies, clinics, or other such institutions and facilities; It can be provided indirectly to the patient, in which case the clear solution can be up to 1 liter or more. The larger container may have a larger capacity, and at least one smaller amount of antibacterial agent may be dispensed from the larger container. The solution is withdrawn one or more times and transferred to a smaller vial and dispensed by the pharmacy or clinic. can be provided to customers and / or patients.

[0144] A single vial system that includes a pen-type injector device for delivering the solution. Approved devices include BD Pens, BD Autojector®, , Humaject®, NovoPen®, BD®P en, AutoPen®, and OptiPen®, Genotro pinPen(R), Genotronorm Pen(R), Humat r ○ Pen (registered trademark), Reco-Pen (registered trademark), Roferon Pen ( Registered trademark), Biojector (registered trademark), Iject (registered trademark), J-tip Needle-Free Injector (registered trademark), Intraject (registered trademark) trademark), Medi-Ject (registered trademark), Smartject (registered trademark) (e.g., B ecton Dickensen(Franklin Lakes, NJ, www.b ectondickenson.com), Disetronic (Burgdorf, Switzerland, www.disetronic.com;)Bioject, Portland,Oregon(www.bioject.com), Nationa l Medical Products,Weston Medical(Peterb orough, UK, www.weston-medical.com), Medi-J ect Corp(Minneapolis,MN,www.mediject.com ) and similar suitable devices. Approved devices that contain dual vial systems include the HumatroPen® Reconstituting the lyophilized drug in a cartridge for delivery of the reconstituted solution, such as Other suitable devices include pen-type injector systems for administering the drug to the patient. These include pre-filled syringes, auto-injectors, needleless syringes, and needleless IV infusion sets. can be done.

[0145] The product may include packaging material that, in addition to any information required by regulatory authorities, The packaging material of the present invention, if applicable, provides at least Each anti-IL-23 antibody is reconstituted in an aqueous diluent to form a solution, and incubated for 2 to 24 hours or more. The instructions were to use this solution for two vials of wet / dry products over a period of time. Provide to patient. In the case of single vial solution products, prefilled syringes, or auto-injectors, In such cases, the label shall indicate that such solution can be used for a period of 2 to 24 hours or more. The product is useful for human pharmaceutical product applications.

[0146] The formulations used in the methods of the invention comprise an anti-IL-23 antibody and a selected buffer, preferably The process includes mixing saline or a phosphate buffer containing a selected salt. The anti-IL-23 antibody and the buffer are mixed in an aqueous diluent, Conventional dissolution and mixing procedures are used to prepare suitable formulations, for example: A quantity of at least one antibody in water or buffer is added to the desired concentration of protein and buffer. and the desired buffer in a sufficient amount of water to provide. For example, the order of addition of the components, the amount of additional additives, etc., will be recognized by those skilled in the art. Whether or not it is used, the temperature and pH during preparation of the formulation, and the administration concentration and means used It is a factor that can be optimized.

[0147] The methods of the present invention include a variety of formulations that are useful and acceptable for administration to a human or animal patient. The pharmaceutical composition includes water at "normal" conditions as a diluent, and They are prepared using routine methods well known to those skilled in the art. For example, histidine and histidine- The buffer components, such as hydrochloride hydrate, are provided first, followed by the appropriate non-final volumes of “standard” A water diluent, sucrose, and polysorbate 80 may then be added. Finally, the antibody can be added in a "standard" condition using water as the diluent. The volume of the pharmaceutical composition is adjusted to the desired final volume under the above conditions. We recognize several other methods that may be suitable.

[0148] The pharmaceutical compositions contain the indicated masses of each component per volume unit of water or are "standardized" in the The term "aqueous solution" as used herein may be an aqueous solution or suspension having a pH value indicated in the "state". The term "standard conditions" means a temperature of 25°C ± 2°C and a pressure of 1 atmosphere. The term "standard conditions" is used in the art to refer to a single temperature or It is not used to refer to a set of pressures, but instead refers to a particular set of pressures under "standard state" conditions. A reference condition that specifies the temperature and pressure used to describe a solution or suspension containing This is because the volume of the solution is in part a function of temperature and pressure. It is recognized that pharmaceutical compositions equivalent to those disclosed herein may be prepared at other temperatures and pressures. Such pharmaceutical compositions are equivalent to those disclosed herein as defined above. should be determined under "standard state" conditions (e.g., 25°C ± 2°C and 1 atmosphere pressure). do.

[0149] Importantly, such pharmaceutical compositions contain "approximately" 100 mg of 10 ... A compound containing a specific value of component mass (e.g., "about 0.53 mg of L-histidine") The pH value of the pharmaceutical composition may be about 100% or about 150%. The pH value may be adjusted to determine whether the isolated antibody is present in a pharmaceutical composition or whether the isolated antibody is in a pharmaceutical composition. After removal from the pharmaceutical composition (e.g., by dilution), the isolated antibody present in the pharmaceutical composition is The given number is "about" the number of times that the component can be bound to a peptide chain. The mass value or pH value of the isolated antibody after placing the isolated antibody in a pharmaceutical composition. A given value is "about" when the binding activity of the antibody is maintained and detectable.

[0150] Competitive binding assays were performed to determine whether IL-23-specific mAbs bind to similar or different epitopes. Determine whether Abs bind and / or compete with each other. Competing mAb is added, followed by biotinylated hrIL-23. For positive controls, the same mAb was used as a competitor mAb ("self-competition") for coating. IL-23 binding may be detected using streptavidin. The results will show whether the mAbs recognize similar or overlapping epitopes on IL-23. Indicates whether

[0151] One aspect of the methods of the invention involves administering a pharmaceutical composition to a patient.

[0152] In one embodiment of the pharmaceutical composition, the isolated antibody concentration is about 7 mL per mL of pharmaceutical composition. In another embodiment of the pharmaceutical composition, the pH is about 5.5 to about 6.5. be.

[0153] Stable or preserved formulations may be formulated as clear solutions or in aqueous diluents without preservatives or buffers and At least one lyophilized anti-I antibody, which is reconstituted in a second vial containing an excipient. It may be provided to the patient as a dual vial containing a vial of the L-23 antibody. Either single solution vials or dual vials requiring reconstitution are reused multiple times. can be used to fulfill single or multiple patient treatment cycles and therefore are This provides a more convenient treatment regimen than is currently available.

[0154] Other formulations or methods for stabilizing anti-IL-23 antibodies include permeation of lyophilized powders containing the antibodies. The solution may be other than a clear solution. Non-clear solutions include formulations that contain particulate suspensions. Such particles may be microspheres, microparticles, nanoparticles, nanospheres, or liposomes. Compositions containing anti-IL-23 antibodies in structures of various sizes known variously as globulins. Such relatively homogeneous essentially spherical microparticle formulations containing an active agent are disclosed in U.S. Pat. As taught in US Pat. No. 4,589,330, an aqueous phase containing an active agent and a polymer and a non-aqueous phase and then evaporating the non-aqueous phase to cause coalescence of the particles from the aqueous phase. Porous microparticles can be formed as taught in U.S. Pat. No. 4,818,542. As described above, a first phase containing an active agent and a polymer dispersed in a continuous solvent is used, and a frozen It can be prepared by removing this solvent from the suspension by drying or dilution-extraction-precipitation. Preferred polymers for such preparations are gelatin agar, starch, arabinogalactam, Albumin, collagen, polyglycolic acid, polylactic acid, glycolide-L(-) lactate Poly(epsilon-caprolactone), Poly(epsilon-caprolactone-co-lactide acid), Poly(epsilon-caprolactone-co-glycolic acid), Poly(β-hydroxy Poly(alkyl-2-cyanoacrylate), Polyethylene oxide, Polyethylene, Poly(alkyl-2-cyanoacrylate) Poly(hydroxyethyl methacrylate), polyamide, poly(amino acid), poly Poly(2-hydroxyethyl DL-aspartamide), poly(ester urea), poly(L -phenylalanine / ethylene glycol / 1,6-diisocyanatohexane) and poly (methyl methacrylate), Particularly preferred polymers are polyglycolic acid, polylactic acid, glycolide-L (-) Lactide Poly(epsilon-caprolactone), Poly(epsilon-caprolactone) Poly(epsilon-co-lactic acid), and poly(epsilon-caprolactone-co-glycolic acid), etc. It is a polyester of the formula: Water, hexafluoroisopropanol, methylene chloride, tetrahydrofuran, hexane , benzene, or hexafluoroacetone sesquihydrate. The active substance-containing phase is called the second phase. The process of dispersing the first phase into the second phase is done by forcing the first phase through an orifice in a nozzle using pressure. The step of influencing droplet formation may be included.

[0155] Dry powder formulations can be prepared, for example, by spray drying, or solvent extraction by evaporation, or aqueous or Solvent extraction by precipitation of the crystalline composition followed by one or more steps to remove the non-aqueous solvent. The antibody formulation may also be obtained as a result of processes other than lyophilization, such as spray drying. Antibody-based dry powder compositions are taught in U.S. Patent No. 6,019,968. under conditions to provide a solution or slurry of the body, and optionally, a respirable dry powder. The excipients in the solvent can be produced by spray drying. The stability of antibodies is affected by oxygen-inhibited compounds, such as polar compounds like water and ethanol. The spray drying procedure may be carried out under, for example, a nitrogen blanket, or in the presence of a drying gas. This can be augmented by using nitrogen as a suction source. Another relatively dry formulation is Typically, hydrofluoroalkanes such as those taught in WO 9916419 are used. The stabilized dispersion is a dispersion of a plurality of porous microstructures dispersed in a suspension medium containing a propellant. The resulting dispersion can be administered to the patient's lungs using a metered dose inhaler. Equipment useful in the manufacture of fluororesin is manufactured by Buchi Ltd. or Nir ○ Corp. It is being done.

[0156] The anti-IL-23 antibodies in any of the stable or preserved formulations or solutions described herein As is well known in the art, the present invention can be administered by SC or IM injection, percutaneous, pulmonary, mucosal, or implantation. The pump may be an osmotic pump, a cartridge, a micropump or any other pump known in the art. The subject matter of the present invention can be delivered via a variety of delivery methods, including other means that will be understood by those of skill in the art. can be administered.

[0157] therapeutic application The present invention also provides a method for the preparation of ... medicament for the treatment of a brain tumor comprising administering to the patient a therapeutically effective amount of at least one of the following: The IL-23 antibodies of the present invention can be used to, for example, in a cell, tissue, organ, animal, or patient. A therapeutically effective amount of an IL-23 specific antibody is administered or contacted to a cell, tissue, organ, animal, or The present invention also provides a method for modulating or treating psoriasis in a patient.

[0158] Any of the methods of the invention may be used to treat a cell, tissue, or organ in need of such modulation, treatment, or therapy. administering to the organ, animal, or patient an effective amount of a composition or pharmaceutical composition comprising an anti-IL-23 antibody; Such methods may optionally include administering to a subject a subject for the treatment of such a disease or disorder. The co-administration or combination therapy may further include administering to a subject the at least one anti-IL-2 3. Administering the antibody, specified portion, or variant thereof may be in combination with at least one TNF antagonist ( For example, but not limited to, chemical or protein TNF antagonists, TNF monoclonal antibodies, monoclonal or polyclonal antibodies or fragments, soluble TNF receptors (e.g., p5 5, p70, or p85) or fragments, fusion polypeptides thereof, or small molecule TNF antagonists Drugs such as TNF binding protein I or II (TBP-1 or TBP-II), Nereli Monomab, infliximab, etanercept (Enbrel™) , adalimumab (Humira™), CDP-571, CDP-870, Afelix tumour, lenercept, etc.), antirheumatic drugs (e.g. methotrexate, auranofil, aurothioglucose, azathioprine, sodium gold thiomalate, hydroxysulphate cyclophosphamide, leflunomide, sulfasalazine), muscle relaxants, narcotics, nonsteroidal antidepressants Inflammatory drugs (non-steroid anti-inflammatory drugs, NSAIDs), analgesics, anesthetics, sedatives , local anesthetics, neuromuscular blocking agents, antibacterials (e.g., aminoglycosides, antifungals, antiparasitics Drugs, antivirals, carbapenems, cephalosporins, fluoroquinolones, macrolides , penicillins, sulfonamides, tetracyclines, and other antibacterial drugs), psoriasis medications, Tychosteroids, anabolic steroids, diabetes-related drugs, minerals, nutritional drugs, thyroid drugs , vitamins, calcium-related hormones, antidiarrheals, antitussives, antiemetics, antitumor drugs, laxatives, anti Coagulants, erythropoietin (e.g., epoetin alfa), filgrastim (e.g., , G-CSF, Neupogen), Sargramostim (GM-CSF, Leukine ), immunosuppressants, immunoglobulins, immunosuppressants (e.g., basiliximab, cyclosporine daclizumab), growth hormone, hormone replacement drugs, estrogen receptor modulators, mydriasis agents, cycloplegics, alkylating agents, antimetabolites, mitotic inhibitors, radiopharmaceuticals, antidepressants , antimanic drugs, antipsychotic drugs, antianxiety drugs, hypnotics, sympathomimetics, stimulants, donepezil, tac Phosphorus, asthma medications, beta agonists, inhaled steroids, leukotriene inhibitors, methyl Xanthines, cromolyn, epinephrine or analogs, dornase alfa (Pulmozym e) at least one selected from a cytokine or a cytokine antagonist; Suitable dosages can be determined by methods known in the art. For example, Wells et al., eds., Pharmacoth erapy Handbook, 2 nd Edition, Appleton and Lange, Stamford, CT (2000), PDR Pharmacopoei. a, Tarascon Pocket Pharmacopoeia 2000, Del uxe Edition, Tarascon Publishing, Loma Lin da, CA (2000), Nursing 2001 Handbook of Dru. gs,21 st edition,Springhouse Corp.,Spring house, PA, 2001, Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Pre. ntice-Hall, Inc., Upper Saddle River, NJ. and each of these references is incorporated herein by reference in its entirety.

[0159] Treatment Typically, treatment of psoriasis will take approximately 10 minutes, on average, depending on the specific activity of the active agent contained in the composition. A total of at least about 0.01 to 500 milligrams per kilogram of patient body weight per dose. and preferably at least 100 mg / kg of the patient's body weight per single or multiple dose. Anti-IL-23 antibody compositions in the range of at least about 0.1 to 100 milligrams of antibody per kg. The effective serum concentration is affected by administering an effective amount or dose of the composition. In one or more doses, the serum concentration may be from 0.1 to 5000 μg / mL. Dosages will be known to medical practitioners and will, of course, depend on the particular disease state, the Depending on the specific activity of the composition being treated, and the particular patient being treated, Repeated administration, i.e., repeated doses of specific monitored or fixed amounts, may be used to obtain a desired therapeutic dose. It may be necessary to provide separate doses, in which case the separate doses may be used to determine the desired daily dose. Or repeated until an effect is achieved.

[0160] 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, 1 3, 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, 5 3, 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, 9 4, 95, 96, 97, 98, 99 and / or 100-500 mg / kg / dose, or includes any range, value, or fraction thereof, or 0 per single or multiple dose. .1, 0.5, 0.9, 1.0, 1.1, 1.2, 1.5, 1.9, 2.0, 2.5, 2 .9, 3.0, 3.5, 3.9, 4.0, 4.5, 4.9, 5.0, 5.5, 5.9, 6 .0, 6.5, 6.9, 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9 .5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 20, 12.5 , 12.9, 13.0, 13.5, 13.9, 14.0, 14.5, 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, 2 2, 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 of serum The concentration may be included to obtain a mid-level concentration, or any range, value, or fraction thereof.

[0161] Alternatively, the dose to be administered may be determined based on the pharmacodynamic characteristics of the particular agent and its method and route of administration. , age, health and weight of the recipient, nature and severity of symptoms, type of concurrent treatment, frequency of treatment The dosage of the active ingredient may vary depending on known factors such as the dosage, the amount of the active ingredient administered, and the desired effect. It may be about 0.1 to 100 milligrams per kilogram of weight. Usually, it is 0.1 to 50, preferably Preferably, 0.1 to 10 milligrams per kilogram per dose, or in sustained release form, is used to obtain desirable results. It is effective to obtain

[0162] As a non-limiting example, human or animal treatment may be performed using a single dose, an intravenous dose, or multiple doses. Use 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, 2 9th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th or 40th day On at least one of these days or in addition, 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, 3 6, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or in at least one of weeks 50, 51, or 52, or in addition, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, At least one of the 18th, 19th, or 20th years, or any combination of these 0.1 to 100 mg / kg per day, e.g., 0.5, 0.9, 1.0, 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 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 the composition of the present invention. It may be provided as a single or periodic dose of the at least one antibody.

[0163] Dosage forms (compositions) suitable for internal administration generally contain about 0.001 milligrams per unit or container. These pharmaceutical compositions contain from about 1000 mg to about 500 milligrams of active ingredient. is typically present in an amount of about 0.5 to 99.999% by weight, based on the total weight of the composition.

[0164] For parenteral administration, the antibody may be administered either together with a pharma- ceutically acceptable parenteral vehicle or separately. provided as a solution, suspension, emulsion, particle, powder, or lyophilized powder, Examples of such vehicles include water, saline, Ringer's solution, dextrose, etc. Solutions and 1-10% human serum albumin. Non-aqueous solutions such as liposomes and fixed oils. A vehicle may also be used. The vehicle or lyophilized powder may be isotonic and chemically stable. Additives to maintain isotonicity (e.g., sodium chloride, mannitol; chemical stability) The formulation may contain buffers and preservatives (e.g., for stability). The device is surgically sterilized.

[0165] Suitable pharmaceutical carriers are described in Remington's, a standard reference text in this field. Described in the latest edition of Pharmaceutical Sciences, A. Osol It has been done.

[0166] Alternative Administration In order to administer a pharma- ceutical effective amount of an anti-IL-23 antibody, in accordance with the present invention, a number of known The previously described and developed methods can be used. In the following description, pulmonary administration is used. However, other modes of administration may be used in accordance with the present invention with suitable results. The -23 specific antibody is present in a carrier as a solution, emulsion, colloid or suspension. or as a dry powder, by inhalation or as described herein or in the art. 2. The method of claim 1, further comprising administering the composition of the present invention to a subject using any of a variety of devices and methods suitable for administration by other methods known in the art. and deliver it.

[0167] Parenteral Formulation and Administration Formulations for parenteral administration contain sterile water or saline, polyethylene glycol, and glycerol as common excipients. Even if it contains polyalkylene glycols such as ethanol, vegetable oils, hydrogenated naphthalene, etc. Aqueous or oily suspensions for injection may be prepared by adding suitable emulsifying agents or wetting agents and Injections can be prepared, for example, by using aqueous solutions, sterile injectable solutions, and suspensions. The diluent may be a non-toxic parenterally administrable diluent such as a suspension in a solvent. The vehicle or solvent may be water, Ringer's solution, isotonic saline, or the like. As a solvent or suspending medium, sterile, fixed oils can be used. , natural or synthetic or semi-synthetic, fatty oils or fatty acids, natural or synthetic or semi-synthetic, Any type of fixed oil, including monoglycerides, diglycerides, or triglycerides Parenteral administration is well known in the art and can be used in the art. A conventional injection device, such as a gas-pressurized needleless injection device as described in U.S. Pat. No. 5,851,198. injection devices, and laser perforation devices as described in U.S. Pat. No. 5,839,446. These include, but are not limited to, drilling devices, all of which are incorporated herein by reference in their entirety. is incorporated into the book.

[0168] alternative delivery The present invention further relates to parenteral, subcutaneous, intramuscular, intravenous, intraarticular, intrabronchial, intraperitoneal, intracapsular, intrachondral, Intracavitary, intracavity, intracerebellum, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic , intrapericardium, intraperitoneum, intrapleura, intraprostate, intrapulmonary, intrarectum, intrakidney, intraretina, intraspinal cord, bursa Intrathoracic, intrauterine, intravesical, intralesional, bolus, intravaginal, rectal, oral, sublingual, intranasal, The anti-IL-23 antibody composition is administered parenterally or via a transdermal route. Oral (subcutaneous, intramuscular or intravenous) or any other administration, particularly liquid solutions or suspensions In particular, semi-prepared formulations, such as, but not limited to, creams and suppositories, are suitable for use in the form of In solid form, such as tablets or capsules, for use in vaginal or rectal administration. For oral or sublingual administration, or in the form of powder, nasal spray, etc., but not limited to or in the form of, but not limited to, an aerosol or certain medications, either intracavity, or by modifying the skin structure, or by increasing drug concentration in transdermal patches. For either, chemical promoters such as dimethyl sulfoxide (Junging er, et al. In “Drug Permeation Enhancement; "Hsieh, DS, Eds., pp.59-90 (Marcel Dekker, Inc. New York 1994, which is incorporated herein by reference in its entirety), The application of protein and peptide-containing formulations to the skin (WO 98 / 53847 No. 5, or for creating a transient transport pathway, such as by electroporation, or Application of an electric field to increase the mobility of a charged drug through the skin, such as in iontophoresis or application of ultrasound, such as sonophoresis (U.S. Pat. Nos. 4,309,989 and 4,713,621). No. 67,402) by using an oxidizing agent that enables the preparation of a gel, ointment, lotion, suspension or The compositions may be prepared transdermally, such as, but not limited to, a patch delivery system. (The above publications and patents are incorporated herein by reference in their entireties.)

[0169] Methods of sales and / or promotion The present invention further relates to antibodies against IL-23, such as the antibodies described herein (e.g., Approved drugs (approved by the U.S. FDA or equivalent export regulatory agency) The present invention relates to a method for marketing and / or promoting guselkumab (Tre mfya®) in connection with the launch of psoriasis treatment with an antibody against IL-23 Clinical endpoint results at 44 and / or 48 weeks after initial treatment in patients Results are as follows: After continuous treatment with secukinumab, at 44 and / or 48 weeks after initial treatment Advertising, promotion and / or advertising of the superiority of the clinical endpoint results over those of includes highlighting in a different way.

[0170] Having generally described the present invention, the same is provided by way of illustration and not limitation. This will be more readily understood by reference to the following examples, which are not intended to be exhaustive. Further details of the present invention are illustrated by the following non-limiting examples. The disclosures of all citations are expressly incorporated herein by reference.

[0171] Example 1: CNT ○1959 (Gsellschaft für der Staphylococcus aureus) for the treatment of moderate to severe plaque psoriasis A multicenter, randomized, double-blind, randomized, double-blind study to evaluate the comparative efficacy of cumab and secukinumab Double-blind Phase 3 clinical trial Experimental Design: - Two concurrent treatment arms (guselkumab 100mg and secukinumab 300mg) A multicenter, randomized, randomized study in subjects with moderate to severe plaque psoriasis A double-blind, Phase 3, active comparator controlled trial. Randomization: At week 0, approximately 1040 subjects who met all inclusion and exclusion criteria were randomized to Subjects were randomized to 1: They were planned to be randomized to one of two groups in a 1:1 ratio: Group I (n=520): Weeks 0, 4, 12, and 20 From the first week onwards, guselkumab 100 mg was administered subcutaneously every 8 weeks until the 44th week. Group II (n=520): Weeks 0, 1, 2, 3, and 4, and From week 4 onwards, secukinumab 300 mg was administered subcutaneously every 4 weeks until week 44. Treatment period / trial period: Week 44 was the last visit for administration. Week 44 and after Participants were followed for an additional 12 weeks with a final safety visit at week 56. The study was to end when the last subject completed the 56-week visit. So, as of week 56, we had one database lock (DBL).

[0172] Table 4 below provides an overview of the clinical trial.

[0173] [Table 2] Primary analysis set for efficacy: The primary efficacy analysis included all patients with ≥ 18% or more of the treatment received, regardless of the treatment actually received. All randomized subjects were randomized according to their assigned treatment at week 0. This is also called the complete analysis set (FAS). The complete analysis set includes all It was also used in the secondary efficacy analysis. Primary endpoint: Proportion of subjects achieving PASI90 response at 48 weeks (non-inferiority) trial, followed by a superiority trial) Key secondary efficacy variables: There were six key secondary endpoints in this study. There was. The proportion of subjects who achieved PASI 75 response at both 12 and 48 weeks (non-inferiority trial, followed by a superiority trial) The proportion of subjects who achieved a PASI90 response at week 12 (non-inferiority study) Proportion of subjects who achieved PASI 75 response at week 12 (non-inferiority study) Proportion of subjects achieving a PASI 100 response at week 48 (non-inferiority trial, followed by superiority trial) Sexuality Test) Proportion of subjects achieving an IGA score of cure (0) at Week 48 (non-inferiority study, followed by (superiority trial) The proportion of subjects who achieved a cure (0) or minimal (1) IGA score at Week 48 (non- Inferiority trials followed by superiority trials) Non-inferiority margins were set at 10% for all endpoints. To control for the overall type 1 error rate, the primary and main secondary analyses were It was specified that the tests would be performed in a fixed order as indicated. The secondary endpoints were tested only if the primary endpoint was positive and were followed up An endpoint is tested only if the preceding endpoint is positive. I made it so that. Planned sample size and power: Approximately 1,040 total subjects randomized in a 1:1 ratio Human subjects were randomly assigned to receive the 48-week randomized controlled trial with at least 92% power and a two-sided significance level of 0.05. In the same study, the PASI90 response rate was significantly higher in the guselkumab and secukinumab groups. It was expected that the study would detect differences between the two groups. 1 and CNTO1959PSO3002 and secukinumab Phase 3 clinical trial (ERAS Based on the data from the URE and FIXTURE, sample size and power calculations were performed. The assumption is, The PASI90 response rate at week 48 was 70% to 80% in the guselkumab group. % was achieved in the primary endpoint, and 60% to 70% in the secukinumab group.

[0174] Based on the above assumptions, the planned sample size, and a 10% non-inferiority margin, The power to demonstrate non-inferiority for the primary endpoint of PASI90 at week 8 was: It was thought to be greater than 99%.

[0175] Primary Objectives: The primary objective is to evaluate Guselkumab for the treatment of subjects with moderate to severe plaque psoriasis. The aim of this study was to evaluate the efficacy of secukinumab compared with nivolumab.

[0176] Most important results summary CNTO1959PSO3009 is IGA≧3 and PASI≧12. and psoriasis affecting at least 10% of the body surface area (BSA involvement). Potential or actual prior systemic or phototherapy treatment A multicenter, randomized controlled trial in subjects with moderate to severe plaque psoriasis, as defined by This is a randomized, double-blind, phase 3, controlled trial. The database lock was performed for all randomized subjects through week 56. Includes data on.

[0177] A total of 1200 subjects were screened, of which 1048 were selected for Guselk Patients were randomized to receive either secukinumab (n=534) or secukinumab (n=514). The experiment was conducted in nine countries (Australia, Canada, Czech Republic, France, Germany, Hungary, Poland, The study was conducted at 141 sites in the United States, Canada, and the United States. Treatment groups were: baseline population The majority of subjects were Caucasian (93.6%) and had a psoriasis profile. The median age was 46.0 years and the mean age was 18.4 years. The baseline weight was 89.2 kg (Appendix 1). Subjects did not receive any of the study medications due to violations of the study entry criteria. Three subjects were included in all efficacy analyses but excluded from safety analyses.

[0178] Baseline disease characteristics were generally similar between treatment groups. Median duration of psoriasis was 16. The median percentage of body surface area (BSA) affected by psoriasis was 20.0 The median PASI score was 18.0. In addition, 76.1% of subjects had an I GA=3, and 23.8% of subjects had a baseline IGA score of 4. Patients had severe disease, as defined by the WHO Guidelines for Clinical Trials (Appendix 2).

[0179] The proportion of subjects previously treated with each previous psoriasis medication category was Overall, 51.8% had previously received phototherapy and 53.7% had previously received all-cause 1% had received prior biologic therapy, and 29.1% had received prior biologic therapy. Overall, 30.0% of subjects 7.1% had never received either non-biologic or biologic systemic therapy (Appendix 3).

[0180] Key baseline demographics, psoriasis disease characteristics, and prior psoriasis medications / therapies were presented. To summarize in 1.

[0181] [Table 3] By week 44, 5.1% of subjects in the guselkumab group and 9.0% of subjects in the secukinumab group had The most common reasons for discontinuing study medication were: The incidence of adverse events (1.7%) and subject withdrawals (1.3%) in the guselkumab group was %), and in the secukinumab group, subjects withdrew from the study (3.7%) and experienced adverse events (2. 1%) (Appendix 4).

[0182] Primary Efficacy Endpoints: At week 48, 70.0% of subjects in the secukinumab group achieved a PASI90 response. The proportion of patients with refractory glaucoma in the guselkumab group was significantly higher (84.5%) than in the control group (p < 0. 001) (Table 2).

[0183] [Table 4]

[0184] Key secondary efficacy endpoints: - Guselkumab demonstrated a statistically significant improvement in the proportion of subjects who achieved a PASI 75 response at Week 12 and Non-inferiority at both time points at 48 weeks [84.6% (guselkumab) vs. 80.2% (selenium) cukinumab), 95% CI: (-0.2%, 8.9%), p<0.001] (Appendix 6 ), however, the response rate in the guselkumab group was numerically lower than that in the secukinumab group. However, the superiority test did not show any significant difference (p=0.062) (Appendix 7). Therefore, the primary and main secondary analyses were performed to control for the overall type 1 error rate. As these trials are to be conducted in a fixed order for the remainder of the key secondary endpoints, Therefore, the reported p-values ​​are considered nominal. - Non-inferiority was observed in the proportion of subjects who achieved PASI90 response at 12 weeks was not demonstrated [69.1% (guselkumab) vs. 76.1% (secukinumab), 95% confidence interval: (-12.2%, -1.7%), p=0.127] (Appendix 8). When assessed by the proportion of subjects who achieved a PASI 75 response at week 12, Guserku Mab was non-inferior to secukinumab [89.3% (guselkumab) vs 91.6% ( secukinumab), 95% CI: (-6.0%, 1.2%), p<0.001] (Appendix 9). The proportion of subjects who achieved a PASI 100 response at week 48 was The incidence rate was significantly higher in the secukinumab group than in the guselkumab group [58.2% (guselkumab) vs. 48.4% (secukinumab)]. (secukinumab), p=0.001)] (Appendix 10). The proportion of subjects who achieved an IGA score of cure (0) at week 48 was The rate was significantly higher in the secukinumab group than in the guselkumab group [62.2% (guselkumab) vs. 50 .4% (secukinumab), p<0.001] (Appendix 11). The proportion of subjects who achieved a cure (0) or minimal (1) IGA score at Week 48 was , was significantly higher in the guselkumab group than in the secukinumab group [85.0% (guselkumab mab) vs 74.9% (secukinumab), p<0.001] (Appendix 12).

[0185] Other efficacy endpoints: Patients who achieved PASI90 response at all 7 visits from Week 24 to Week 48 The proportion of patients with refractory pulmonary tuberculosis was significantly higher in the guselkumab group than in the secukinumab group [71. 0% (guselkumab) vs. 61.5% (secukinumab), p<0.001] (Appendix 13). IGA and PASI responses over time

[0186] From week 1 to week 48, the following responses were recorded: PASI90, PASI100, and cure (0). The GA score and the proportion of subjects achieving a cure (0) or minimal (1) IGA score over time. The changes over time are summarised in Figure 1 below (see also Appendices 14 and 15).

[0187] These curves show the rate and duration of responses between guselkumab and secukinumab. The differences in retention are striking. The PASI90 figure panels, for example, show that at both weeks 2 and 3 This indicates that the patient is beginning to respond to the treatment. Secukinumab has a higher PASI90 response rate than guselkumab.1 At weeks 6 and 20, both drugs demonstrated similar PASI90 response rates, with guselkumab The PASI90 response rate for serogroup A was 1.0% at all visits from Week 24 to Week 48. The PASI90 response rate for guselkumab is higher than that for cukinumab. The line reached a plateau at week 28, after which response rates remained stable through week 48. In contrast, the PASI90 response rate curve for secukinumab begins earlier at week 20. A plateau was reached at week 20, after which response rates steadily declined from week 20 to week 48. For the other three endpoints, the timing at which response rates reach a plateau Secukinumab showed a higher response rate than guselkumab. Although there was some variability in the time of visit when the response rate switched to PAS, the response rate pattern was consistent with that of PAS. It is similar to the I90 pattern.

[0188] Safety: Regardless of the treatment assigned at random, a small number of patients were randomized according to the actual treatment they received during the trial. All randomized, randomized, and randomized participants who received at least one dose of study medication (partial or complete) Safety was assessed among all subjects who received treatment with the drug. This is also called the safety analysis set. Key safety events are summarized in Table 3.

[0189] [Table 5] Subjects experiencing one or more adverse events classified by the investigator as an infection The rate was lower in the guselkumab group compared with the secukinumab group ( in the secukinumab group, 58.6% [313 / 534], and in the secukinumab group, 64.8% [ 331 / 511] (Appendix 19). The most common infection was nasopharyngitis (21.9% (guselkumab) vs. 24.5% (seroquel)). cukinumab)] and upper respiratory tract infections [15.5% (guselkumab) vs. 18.0% (secukinumab) The patient was a PT for [Mab]. PTs representing fungal infections reported in >2% of subjects included tinea pedis (guselkumab group 1) .1% in the guselkumab group vs. 3.1% in the secukinumab group), oral candidiasis (0.9% in the guselkumab group vs. 3.1% in the secukinumab group), secukinumab group (2.2%), and vulvovaginal candidiasis (0.9% in the guselkumab group vs. 2.0% in the secukinumab group). .5%) was included. Serious infections were single events in both treatment groups, with no serious infections reported in either treatment group. No cases of active tuberculosis or opportunistic infections were reported during the study. (Appendix 21). A total of 3 cases of basal cell carcinoma (BCC) (0.6%) were reported in the guselkumab arm. Two cases of BCC (0.4%) were reported in the secukinumab group. Two cases of cutaneous squamous cell carcinoma and one case of Bowen's disease were reported in the guselkumab arm. . One subject in the guselkumab arm was diagnosed with invasive ductal carcinoma. One subject was diagnosed with non-small cell lung cancer and one subject was diagnosed with mycosis fungoides. Ta. A total of three subjects in the secukinumab group experienced Crohn's disease, IBD, or colitis events. and reported: ■ One subject was diagnosed with a serious adverse event (AE) due to Crohn's disease. The patients received the study drug as scheduled on five occasions. ■Two subjects reported non-serious AEs for IBD. One subject had a history of chronic IBD that was not identified during screening. had been randomized to receive five doses of the study drug, but subsequently developed colon clotting Treatment was discontinued because chronic rheumatoid arthritis was confirmed.

[0190] The second subject reported symptoms suggestive of Crohn's disease approximately one month after completing 44 weeks of treatment. It was subsequently confirmed that he had Crohn's disease.

[0191] Analysis of patients with psoriatic arthritis (PsA) A subgroup of patients with self-reported infectious arthritis (PsA) was included in the post hoc analysis. Treatment differences and 95% confidence intervals (CIs) were calculated for PsA subgroups. Loss of data was considered non-response. Both efficacy and safety were evaluated up to 56 weeks. Overall, the treatment groups [guselkumab group (GUS) (n=534), secukinumab The subgroup (SEC) (n=514) were comparable at baseline: body weight 89 kg, psoriasis 24% of the total body surface area (PsO) and 24% of the Investigator Global Assessment (IGA). These characteristics were associated with self-reported PsA [GUS( The characteristics of the subgroups with CI (n=97) and SEC (n=79) were similar to those of the overall population. In the whole body, the primary endpoint of PASI90 response achieved at 48 weeks was GUS disease. The first major secondary outcome was 84.5% in patients with SEC vs 70.0% in patients with SEC (P < 0.001). End-point results (achieving PASI75 response at both 12 and 48 weeks) GUS was non-inferior to SEC in 84.6% of patients. However, 80.2% of patients achieved SEC, respectively (p<0.001). No superiority was demonstrated (p=0.062). Among patients with PsA, at 48 weeks Patients who achieved the primary endpoint of PASI90 response at this time point were 82.5% for GUS. %, and 63.3% for SEC (treatment difference 19.2% [95% CI = 5.0, 33 4]. In both the overall population and in the subgroup of patients with PsA, peak PA SI90 response rates were reached between weeks 16 and 24 for both drugs. Patients treated with S maintained this response through week 48, whereas those treated with SEC Patients showed a decreased response rate from weeks 24 to 48. Adverse events observed in the overall population were The safety profile of GUS and SEC was generally consistent with that of other drugs. Safety results among patients were consistent with those in the overall population. In a subset of patients with self-reported PsA, GUS was associated with moderate to severe pulmonary edema. showed greater long-term efficacy and durability of response compared with SEC in the treatment of chronic PsO. However, this was consistent with the overall study population with PsO vulgaris.

[0192] Weight quartile analysis Efficacy data were analyzed by baseline weight quartile (Q1: ≤74 kg, Q2: >74 kg). Q1: 87kg or less, Q2: 87kg or more but less than 100kg, Q3: 87kg or more but less than 100kg, Q4: 100kg or more) and BMI category (normal: 25 kg / m 2 Under and overweight: 25kg / m 2 Over 30kg / m 2 Under 30kg / m 2 In this post-hoc analysis, the baseline Efficacy was assessed by weight quartile and body mass index (BMI) category. There were no weight restrictions at the time of enrollment.

[0193] The data are presented in Tables 12 to 16. After applying the treatment failure rules, missing data were treated as non-response. PA at week 48 in the guselkumab and secukinumab groups, respectively The proportion of patients who achieved a SI90 response was as follows: by baseline weight quartile :Q1: 86.7% vs. 75.6% (11.1% [0.9%-21.3%]), Q2: 89 Q2: 80.3% vs. 71.1% (16.0% [6.0%-26.0%]), Q3: 80.3% vs. 71.1% (16.0% [6.0%-26.0%]), .0% (9.3% [-1.9%-20.6%]), Q4: 82.1% vs. 61.3% (20 .9% [9.4%-32.3%]), by BMI category: 88.1% vs. 75.2% (positive Normal: 12.8% [2.2%-23.5%]), Overweight: 84.1% vs. 73.4% (10. 6% [1.6%-19.7%]), obesity: 82.5% vs. 65.3% (17.2% [8.8 %-25.6%] (percentage difference [95% confidence interval]). These results were consistent with those of all study participants. This was consistent with the primary endpoint of PASI90 at 48 weeks in the group (Grade 1). Selkumab 84.5% vs. secukinumab 70.0% [14.2% (9.2%-19.2%) For PASI100, IGA0, and IGA0 / 1 responses, all weight quartiles Similar results were observed across all BMI categories, with all treatment differences being In conclusion, baseline weight quartile and BMI Across categories, efficacy outcome response rates at 48 weeks were 1. In treatment, guselkumab was consistently and numerically greater compared with secukinumab.

[0194] body region analysis As shown in Table 17, guselkumab showed significant efficacy in treating moderate to severe psoriasis. Compared to cukinumab, body region components including head and neck, trunk, and upper and lower extremities In the control group, PASI showed numerically higher efficacy up to 48 weeks. Head and neck, Improvements in PASI physical domain components, including the trunk, and upper and lower extremities, were also assessed. Missing data were considered as non-response.

[0195] At week 48, guselkumab was associated with a numerically greater proportion of patients compared with secukinumab. The subject performs the PASI components of the head and neck, torso, and upper and lower extremities. ) improvements (100% improvement and ≥90% improvement) were achieved (Table 17).

[0196] Patient geographic region analysis Patients (n=391) from North America (United States, Canada), Eastern Europe (Czech Republic, Hungary) , patients from Poland (n=338), Western Europe (France, Germany, Spain) Patients from the US (n=248) and Australia (n=71) were randomized guselkumab 1 mg / kg at weeks 0, 4, and 12, and every 8 weeks thereafter 00 mg subcutaneously (SC) (n=534), or at weeks 0, 1, 2, 3, Secukinumab 300 mg administered subcutaneously (SC) at week 4 and every 4 weeks thereafter. ) (n=514). Injections were continued through week 44 in both groups. The primary endpoint was and the proportion of patients achieving a PASI90 response at week 48. Everyone agreed.

[0197] As shown in Table 18, at week 48, regardless of geographic region, The PASI90 response rate at time point was In subgroup analyses by geographic region, , patients treated with guselkumab had higher PA compared with patients treated with secukinumab SI90 response rates were: North America (78.9% for guselkumab vs. 60.4% for secukinumab); Eastern Europe (90.6% guselkumab vs. 76.0% secukinumab), Western Europe (Guselkumab vs. Selkumab 82.9% vs. secukinumab 74.8%, and Australia (guselkumab 91.4% vs. 77.8% with secukinumab) (Table 18).

[0198] Example 2: Effects of IL-17F and IL-22 on immune cell populations in skin and serum Evaluation of the therapeutic effects of anti-IL-23 and anti-IL-17A Skin biopsies were collected at weeks 0, 4, and 24. Skin gene expression profiles Isolates were generated in whole biopsies via RNA sequencing. T cell composition was analyzed by flow cytometry. Using quantification in combination with unbiased clustering analysis, Serum was collected at weeks 0, 4, 5, 6, 7, 8, 9, and 10, and was analyzed by immunophenotyping of cell suspensions from the mice. The results were collected at weeks 24 and 48, and IL17A, IL17F, IL22, IL23, and The concentration of β-defensin-2 (BD-2) was analyzed by an ultrasensitive immunoassay. t, CD4 + T cells and CD8 + The number of T cells was measured in the skin lesions.

[0199] result Serum IL17A, IL17F, and IL22 concentrations were measured at 4 and 2 weeks after guselkumab treatment. The incidence of rheumatoid arthritis was reduced at weeks 4 and 48. In contrast, treatment with secukinumab did not improve was less efficient at reducing IL17F concentrations than treatment with lucumab (p<0.0001, at all time points) and had no effect on IL22 concentrations (free I (The L17A concentration could not be measured in the assay used). Compared with treatment with tafamidis, treatment with guselkumab had the same effect at weeks 4, 24, and 48. In the eye, serum concentrations of IL-17F and IL-22 were reduced to a greater extent.

[0200] The concentration of BD-2, a biomarker highly correlated with skin inflammation, decreased at the 4th week was greater with secukinumab compared to guselkumab (p<0.0001), and At week 4, the results were comparable. However, at week 48, BD-2 was superior to Secukinuma. The IL-10 expression level increased in the rituximab group but remained decreased in the guselkumab group (p<0.05). As a result, at week 48, BD-2 concentration with guselkumab was significantly higher than that with secukinumab. The normalization of changes in skin transcription was greater with secukinumab at week 4. The difference in the mean mean mean age at presentation was greater in the guselkumab group than in the IL group, but was comparable at week 24. Normalization of increased skin gene expression of IL-17A, IL-22, and IL-23 at weeks 4 and 24 At week 14, IL23R expression was comparable between treatments, but was significantly higher with guselkumab. At 24 weeks of treatment, CD4 + T cells and CD4 8 + The number of T cells was decreased in the skin lesions in both groups. 8 + TRM(CD3 + , CD8 + , CD103 + , and / or CD49a + ) frequency is There was a decrease in serotonin levels compared to baseline with guselkumab (p=0.036), whereas secukinumab The IL17A expression in the affected skin was not decreased in the control group. + / CD8 + Low TRM The results did not differ between treatments. In contrast, regulatory T cells (Treg) (FoxP3 + , CD25 + ,IL17A - ) was more frequent in the guselkumab group at week 24 was higher (p=0.042).

[0201] Psoriasis translocations that are better normalized by GUS to SECs, including IL23R We identified genes that are part of the scriptome.

[0202] PSO disease of mucosal-associated invariant T cell (MAIT (IL23R included) related genes) Increased expression in the affected skin indicates that GUS is better able to stimulate SECs at 24 weeks. Nicely normalized.

[0203] CD8+ tissue resident memory cells (TRM) (CD3+, CD8+, CD103+, and / or The frequency of CD49a+ or CD49a+ cells was significantly higher in patients treated with GUS than in those treated with ESCC at 24 weeks. Regulatory T cells (Treg) (CD3+, FoxP3+, CD25+, IL17A-) at 24 weeks compared with the SEC group. and was higher in the GUS group (p<0.05).

[0204] Analysis of IL-23+APCs revealed that CD14+CD64+DCs were predominant in PsO lesional skin. It has been shown that CD4 T cell proliferation is responsible for most of the IL-23 expression in non-T This was associated with a relative increase in RM (CD103-, CD49a-). was associated with a relative increase in the frequency of TRM (CD103+ and / or CD49a+). CD8 TRM and non-TRM in CD4+ non-TRM and CD8+ TRM in PsO skin The large increase in CD4 non-TRM and I in PsO (baseline) was The frequency of the Treg population in T cells is a major contributing factor to the progression of PsO lesions. Two distinct clusters of Tregs (one IL17A+ and one IL The T cell subpopulations in lesional versus non-lesional skin were significantly increased. The frequency of IL17A expression in the subset was increased. CD4+ T cells producing IL-17A Most of the CD8+ T cells produced IL-17A, whereas most of the CD8+ T cells produced IL-17A. Most of the Tregs expressed low levels of IL17A in the PsO skin. At week 24, the guselkumab arm showed a In response to guselkumab, the frequency of TRM in CD8+ T cells was more significantly reduced. At week 24, CD4+ T cell No differences in the incidence of TRM were observed among the subsets. Between treatment with cumab and treatment with secukinumab, there was a median CD4+ or No differences were observed in CD8+ T cell subsets in the 14 patients with IL17A. The SEC group was significantly decreased in the control group (measurements in the SEC group indicate that the assay is This complicates matters because it is impossible to distinguish it from IL17A, resulting in an increase in IL17A. The frequency of IL17A expression in CD8 TRMs in lesional skin at baseline The increase in severity was greater than PASI 90 (regardless of treatment group) at week 48 (PASI > 90 In non-lesional skin at baseline, No similar pattern is observed.

[0205] Figure 2 shows the ECLIPSE study design up to week 48 and the biomarker substudy The blood samples for serum protein biomarker subtests are shown. Samples were collected from all enrolled subjects at weeks 0, 4, 24, and 48. At week 0, a pair of skin samples containing non-lesional and lesional skin were collected, and then after 4 weeks and After 24 weeks, skin samples, including lesional skin, were analyzed by skin transcriptome biomarker analysis. A subset of subjects (19 GUS-treated subjects and 16 S subjects) was included for the study. Separately, a pair of non-lesional and lesional skin samples were taken from each subject (EC-treated subjects) at week 0. After 4 weeks and after 24 weeks, skin samples containing lesional skin were A separate subset of subjects (11 subjects) was included for immunophenotyping in the skin immune cell substudy. GUS-treated subjects and 9 SEC-treated subjects).

[0206] As shown in Figure 3, high serum levels of IL-17F in psoriasis patients were associated with both treatments. The reduction was greater and more rapid with guselkumab. Compared with healthy control subjects (n=25), serum levels of IL-17F were significantly higher in patients with psoriasis. The IL-1 level was 5.2-fold higher in subjects (n=200), p<0.0001. The large reduction in serum 7F concentrations was greater with secukinumab at all visits from week 4 onwards. The guselkumab-treated group had a higher mean mean score than the non-guselkumab-treated group (n=100). The mean ... .26 in the secukinumab group compared with 1.12 in the secukinumab group, p<0.0001 and at 24 weeks, 5.32 vs. 2.31, respectively, p<0.00 01, and at 48 weeks, 5.28 times versus 2.33 times, respectively, p<0.01. The mean mean mean score for patients treated with guselkumab at 24 and 48 weeks was .0001. Serum IL-17F concentrations were normalized to those in healthy control subjects. Mean: least squares mean; CI = confidence interval. LS means and 95% confidence intervals are used with repeated measurements. Using a mixed-effects model, the effect of treatment (Guselkumab) was calculated based on log-transformed concentrations. The time points for the study (weeks 0, 4, 24, 48, and 50) and the visits (weeks 0, 4, 24, 48, and 50) were fixed. The outcome is a random effect.

[0207] As shown in Figure 4, high serum levels of IL-22 in psoriasis patients were consistent with both treatments. However, a faster and greater reduction was achieved with guselkumab. Compared to healthy control subjects (n=25), serum levels of IL-22 were significantly higher in psoriasis patients. (n=200), IL-22 was 6.0-fold higher, p<0.0001. The large reduction in serum concentrations of The mean mean mean ≥ 100 was 1.01 at week 4, whereas the mean mean mean ≥ 100 was 1.01 at week 4. The guselkumab group had a 1.74-fold increase in RR and the secukinumab group had a 1.28-fold increase in RR. However, p=0.057, and at 24 weeks, the mean age was 2.79 vs. 1.25, respectively. fold, p<0.0001, and at 48 weeks, 2.85 vs. 1 fold, respectively. .24 times, but p<0.0001.

[0208] LS means: least squares means; CI: confidence interval. LS means and 95% confidence intervals are used to estimate the repeatability of measurements. were calculated based on log-transformed concentrations using a mixed-effects model with Selkumab and secukinumab) and visits (weeks 0, 4, 24, and 48) is a fixed effect and subject is a random effect.

[0209] As shown in Figure 5, elevated levels of β-defensin-2 (BD-2) in psoriasis patients Serum concentrations were reduced by both treatments, but more rapidly with secukinumab. The reduction was more sustained with guselkumab. Serum concentrations of BD-2 were higher in psoriasis patients (n=200) compared with those in the control group (n=25). The reduction in serum BD-2 levels was greater than 32-fold at week 4. At this point, the secukinumab group (n=100) had a significantly higher mean mean score than the guselkumab group (n=100). (13.1 vs. 5.0, p<0.0001), but were comparable at 24 weeks (18.4 vs. 17.3, p=0.99), which was reversed at 48 weeks (Gsell 18.9 times in the cumab group vs. 13.7 times in the secukinumab group, p<0.05).

[0210] LS means: least squares means; CI: confidence interval. LS means and 95% confidence intervals are used to estimate the repeatability of measurements. were calculated based on log-transformed concentrations using a mixed-effects model with Selkumab and secukinumab) and visits (weeks 0, 4, 24, and 48) is a fixed effect and subject is a random effect.

[0211] As shown in Figure 6, a subset of inducible genes in psoriatic lesional skin was In terms of individual differences, guselkumab was better at normalizing the RR-10001-related risk than secukinumab. Quantification of gene expression in skin biopsies was performed using RNA-seq, log2-transformed 1 The mean age of the lesion was 18.1 years and the mean age of the lesion was 18.1 years. Differential gene expression between psoriasis and non-lesional skin (NL) was compared with the corresponding t Based on the test, the ratios were calculated as log2-transformed ratios. 1655 genes were Increased expression in LS with a fold change of >0.5 and a false discovery rate (FDR) of <0.05 Differential gene expression in LS in response to treatment at 4 and 24 weeks was calculated as log2-transformed proportions using a mixed-effects model with repeated measures. , treatment (guselkumab and secukinumab) and visit (weeks 4 and 24) were fixed effects where subject is a random effect and baseline (between lesioned and non-lesional skin) ) are covariates. For a given gene, at a given visit, Percentage improvement in response to treatment was log2-transformed ratio of response in LS versus baseline The mean mean is calculated as the negative ratio of the log2-transformed ratio of the difference between LS and NL at the mean line. Among 55 genes, expression was increased in baseline LS, whereas 328 genes ( 19.8%) responded more significantly to guselkumab than to secukinumab at week 24 The improvement was greater than 50% in response to guselkumab, with was defined as a difference of >25% between guselkumab and secukinumab in The light grey lines represent individual genes, the dark black lines represent the The average improvement rate (%) of 328 genes compared to cukinumab (76%). GUS: Guinea pig Selkumab, SEC: Secukinumab

[0212] As shown in Figure 7, the expression of genes related to mucosa-associated invariant T (MAIT) cells in psoriatic lesions Increased expression in the peripheral skin was greater with guselkumab than with secukinumab at 24 weeks. The light grey lines represent individual genes. GUS: Gus Lucumab, SEC: Secukinumab.

[0213] PsO is a T-cell disorder in which skin-resident T cells produce multiple inflammatory cytokines. , which leads to activation and proliferation of keratinocytes, eventually resulting in excess sebum, erythema, and scaling, P sO is thought to play an important role in regulating the immune response that results in inflammatory features of the skin. Inflammatory T cells in the skin and other tissues are thought to express IL23R and may mediate immune responses. Dependence on IL-23 for pathology. Better understand the mechanism of action of guselkumab. To this end, we performed flow cytometry-based analysis of cell suspensions from dissociated biopsies. We sought to characterize PsO cutaneous T cells by immunophenotyping of the T cells. Using this method, we demonstrated that CD8+ tissue-resident memory in lesional skin at 24 weeks The frequency of T cells (TRM) was decreased relative to baseline levels in the GUS group ( p=0.036), was not reduced in the secukinumab group. At week 24, CD8 had a higher incidence of TRM (p=0.0048).

[0214] [Table 6]

[0215] As shown in Figure 8, at 24 weeks, the TRM of CD8 in PSO skin was The frequency was reduced by guselkumab but not by secukinumab Characterization of psoriatic skin T cells was performed by immunophenotyping cell suspensions obtained from dissociated biopsies. The TRM of CD8 (CD3 + , CD8 + , CD103 + , and / or CD49a + ) was observed in the guselkumab group (n= 11) decreased at 24 weeks (p<0.05), but not in the secukinumab group (n=9). Statistical analysis was performed using SAS 9.4 software using a longitudinal regression model. The difference between the lesions at baseline and the baseline lesions was used as the response. Treatment and week as interaction terms between lesions, factors, and treatment and week in the study, and covariates We used AR1 as the variance structure. We used the least squares method and 95% confidence intervals to estimate the base Data was plotted as change from spine lesion.

[0216] IL23 has also been reported to have antagonistic effects on the function of regulatory T cells (Treg). It has been reported that Tregs increase in the blood and skin of PsO patients. To better understand the mechanism of action of guselkumab, To date, we have performed flow cytometry-based analysis of cell suspensions from dissociated biopsies. We aimed to characterize PsO cutaneous T cells by immunophenotyping. Using this method, Treg (CD3 + , FoxP3 + , CD25 + ) frequency At week 24, the effects were maintained at baseline levels. In the secukinumab-treated group, Treg levels were higher than baseline at week 24. Thus, GUS decreased significantly during the course of treatment compared with T The relative frequency of reg was maintained.

[0217] [Table 7]

[0218] As shown in Figure 9, at 24 weeks, the frequency of regulatory T cells (Treg) was The immunization with numab was reduced, but not with guselkumab. Immunophenotyping of skin cells Characterization of PSO cutaneous T cells by immunization revealed an increase in the frequency of the Treg population compared to baseline. was maintained in the guselkumab group (n=11), but at week 24, 9) showed a decrease (p<0.001). A longitudinal regression model was used and SAS 9. Statistical analysis was performed using software to determine the difference between baseline lesions and response. Predictors of baseline lesions, factors, and interactions between treatment and week Treatment and week as time periods and AR1 as the covariance structure were used. Least squares and Data were plotted as change from baseline lesion size with 95% confidence intervals. Ta.

[0219] To better understand the mechanism of action of guselkumab (GUS) (trade name TREMFYA) To date, we have performed flow cytometry-based analysis of cell suspensions from dissociated biopsies. We sought to characterize PsO cutaneous T cells by immunophenotyping. Using this method, we have demonstrated that CD8+ tissue-resident memory T cells (TRM) in PsO skin The relative frequency of Tregs associated with IL17A was significantly increased by secukinumab, a mAb blocker of IL17A, at 24 weeks. The guselkumab group had a higher The results showed that the association was statistically significant (p=0.006).

[0220] [Table 8]

[0221] As shown in FIG. 10, at 24 weeks, CD8 tissue-resident memory T cells (TR The relative frequency of regulatory T cells (Treg) to secukinumab was significantly higher in the gusel group than in the secukinumab group. Characterization of PSO cutaneous T cells by immunophenotyping of skin cells , Treg population vs. CD8+ compared with secukinumab (n=9) at week 24 The ratio of TRM patients was higher in the guselkumab group (n=11) (p<0.01). Statistical analysis was performed using SAS 9.4 software using a longitudinal regression model. The difference between the lesions at baseline was used as the response, and the baseline lesions as the predictor Treatment and week as interaction terms between lesion, factor, and treatment and week in the covariance structure We used AR1 as the structure. We used the least squares method and 95% confidence intervals to estimate the baseline. Data were plotted as change from lesion area.

[0222] appendix

[0223] [Table 9-1]

[0224] [Table 9-2]

[0225] [Table 10-1]

[0226] [Table 10-2]

[0227] [Table 11-1]

[0228] [Table 11-2]

[0229] [Table 12]

[0230] [Table 13]

[0231] Effectiveness

[0232] [Table 14]

[0233] [Table 15]

[0234] [Table 16]

[0235] [Table 17]

Table 18

[0236]

Table 19

[0237]

Table 20

[0238]

Table 21

[0239]

Table 22-1

[0240]

Table 22-2

[0241]

Table 22-3

[0242]

Table 23-1

[0243]

Table 23-2

[0244] Security

[0245]

Table 24-1

[0246]

Table 24-2

[0247]

Table 24-3

[0248]

Table 24-4

[0249]

Table 24-5

[0250]

Table 24-6

[0251]

Table 24-7

[0252]

Table 24-8

[0253]

Table 24-9

[0254]

Table 24-10

[0255]

Table 24-11

[0256]

Table 24-12

[0257]

Table 24-13

[0258]

Table 24-14

[0259]

Table 24-15

[0260]

Table 25-1

[0261]

Table 25-2

[0262]

Table 25-3

[0263]

Table 26

[0264]

Table 27-1

[0265]

Table 27-2

[0266]

Table 27-3

[0267] [Table 27-4]

[0268] [Table 28-1]

[0269] [Table 28-2]

[0270] [Table 28-3]

[0271] [Table 29]

[0272] [Table 30]

[0273] [Table 31]

[0274] [Table 32]

[0275] [Table 33]

[0276] Summary of data for patients with psoriasis (PsO) and psoriatic arthritis (PsA)

[0277]

Table 34-1

[0278]

Table 34-2

[0279]

Table 34-3

[0280]

Table 35-1

[0281]

Table 35-2

[0282]

Table 36-1

[0283]

Table 36-2

[0284]

Table 37-1

[0285]

Table 37-2

[0286]

Table 37-3

[0287] [Table 38]

[0288] [Table 39-1]

[0289] [Table 39-2]

[0290] [Table 40]

[0291] PASI90 / PASI100 and IGA0 / 1, organized by weight quartiles

[0292] [Table 41]

[0293] [Table 42]

[0294] [Table 43]

[0295] [Table 44] Note: In the secukinumab 300 mg group, 2 patients did not have baseline weight Therefore, only 512 patients were listed in the secukinumab group instead of 514 patients. is.

[0296] [Table 45] Note: There was 1 patient in the guselkumab 100 mg group who did not have baseline height Therefore, in the above analysis, there were 533 patients in the guselkumab group instead of 534 patients. Only patients with baseline height are listed. In the above analysis, 51 patients had baseline height. Because only one patient had a ≥ 300 mg IV schizophrenia, the secukinumab 300 mg group had a ≥ 300 mg IV schizophrenia. Only 511 patients with sline height are listed.

[0297] [Table 46]

[0298] [Table 47]

Claims

1. 1. A method of treating psoriasis in a patient, comprising: administering to said patient an antibody against IL-23 in an amount that has been clinically proven safe and has been clinically proven effective; the antibody against IL-23 comprises a light chain variable region and a heavy chain variable region, The light chain variable region comprises: a light chain complementarity determining region 1 (CDRL1) amino acid sequence of SEQ ID NO:50; The CDRL2 amino acid sequence of SEQ ID NO:56; and a CDRL3 amino acid sequence of SEQ ID NO:73, The heavy chain variable region The heavy chain complementarity determining region 1 (CDRH1) amino acid sequence of SEQ ID NO:5; a CDRH2 amino acid sequence of SEQ ID NO: 20; and a CDRH3 amino acid sequence of SEQ ID NO:44, The patient is treated with the antibody against IL-23 for at least 44 weeks and shows greater efficacy in psoriasis clinical endpoints than patients treated with a secukinumab antibody. The method.

2. 2. The method of claim 1, wherein the psoriasis clinical endpoint is PASI90, PASI100, IGA0, and / or IGA1.

3. The method of claim 2, wherein the psoriasis clinical endpoint is measured 44 and / or 48 weeks after initial treatment with the antibody against IL-23.

4. The method of claim 3, wherein the psoriasis clinical endpoint is measured 48 weeks after initial treatment with the antibody against IL-23.

5. 2. The method of claim 1, wherein the antibody against IL-23 is administered at a first dose, 4 weeks after the first dose, and every 8 weeks after the 4 week dose.

5. 2. The method of claim 1, wherein the secukinumab antibody is administered at a first dose, one week after the first dose, two weeks after the first dose, three weeks after the first dose, four weeks after the first dose, and every four weeks after the fourth week administration.

6. The method of claim 1, wherein the antibody to IL-23 is administered at a dose of 100 mg.

7. 7. The method of claim 6, wherein the antibody to IL-23 is safe and effective for treating psoriasis in areas of the patient selected from the group consisting of the scalp, nails, hands, and feet.

8. the antibody against IL-23 is in a composition, The composition, 100 mg / mL of antibody; 7.9% (w / v) sucrose based on the weight of the pharmaceutical composition; 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate; 0.053% (w / v) polysorbate 80 based on the weight of the pharmaceutical composition; The diluent is water at standard conditions; The method according to claim 6.

9. 10. The method of claim 8, further comprising administering to the patient one or more additional medications used to treat psoriasis.

10. 10. The method of claim 9, wherein the additional agent is selected from the group consisting of an immunosuppressant, a nonsteroidal anti-inflammatory drug (NSAID), methotrexate (MTX), an anti-B cell surface marker antibody, an anti-CD20 antibody, rituximab, a TNF inhibitor, a corticosteroid, and a costimulatory modulator.

11. 2. The method of claim 1, wherein the antibody against IL-23 is effective to reduce symptoms of psoriasis, induce a clinical response, induce or maintain clinical remission, inhibit disease progression, or inhibit complications of disease in the patient.

12. 10. The method of claim 1, wherein the patient is treated for moderate to severe psoriasis.

13. 13. The method of claim 1, further comprising the step of discontinuing treatment of a patient previously treated with at least one dose of secukinumab and deciding to treat said patient with guselkumab.

14. 14. The method of claim 13, further comprising measuring the psoriasis clinical endpoints PASI90, PASI100, IGA0, and / or IGA1 44 and / or 48 weeks after initial treatment, discontinuing treatment of a patient previously treated with at least one dose of secukinumab, and treating said patient with guselkumab.

15. The method of claim 1, wherein the patient is suffering from psoriatic arthritis.

16. 1. A method of treating psoriasis in a patient, comprising administering to said patient an antibody against IL-23 in an amount having clinically proven safety and clinically proven effectiveness; the antibody against IL-23 comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 116 and a heavy chain variable region having the amino acid sequence of SEQ ID NO: 106; The patient is treated with the antibody against IL-23 for at least 44 weeks and shows greater efficacy in psoriasis clinical endpoints than patients treated with a secukinumab antibody. The method.

17. 17. The method of claim 16, wherein the psoriasis clinical endpoint is PASI90, PASI100, IGA0, and / or IGA1.

18. The method of claim 17, wherein the psoriasis clinical endpoint is measured 44 and / or 48 weeks after initial treatment with the antibody against IL-23.

19. The method of claim 18, wherein the psoriasis clinical endpoint is measured 48 weeks after initial treatment with the antibody against IL-23.

20. 17. The method of claim 16, wherein the antibody against IL-23 is administered at a first dose, 4 weeks after the first dose, and every 8 weeks after the 4 week dose.

21. 17. The method of claim 16, wherein the secukinumab antibody is administered at a first dose, one week after the first dose, two weeks after the first dose, three weeks after the first dose, four weeks after the first dose, and every four weeks after the fourth week administration.

22. The method of claim 16, wherein the antibody against IL-23 is administered at a dose of 100 mg.

23. 23. The method of claim 22, wherein the antibody to IL-23 is safe and effective for treating psoriasis in an area of ​​the patient selected from the group consisting of the scalp, nails, hands, and feet.

24. the antibody against IL-23 is in a composition, The composition, 100 mg / mL of antibody; 7.9% (w / v) sucrose based on the weight of the pharmaceutical composition; 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate; 0.053% (w / v) polysorbate 80 based on the weight of the pharmaceutical composition; The diluent is water at standard conditions; 23. The method of claim 22.

25. 25. The method of claim 24, further comprising administering to the patient one or more additional medications used to treat psoriasis.

26. 26. The method of claim 25, wherein the additional agent is selected from the group consisting of an immunosuppressant, a nonsteroidal anti-inflammatory drug (NSAID), methotrexate (MTX), an anti-B cell surface marker antibody, an anti-CD20 antibody, rituximab, a TNF inhibitor, a corticosteroid, and a costimulatory modulator.

27. 17. The method of claim 16, wherein the antibody against IL-23 is effective to reduce symptoms of psoriasis, induce a clinical response, induce or maintain clinical remission, inhibit disease progression, or inhibit complications of disease in the patient.

28. 17. The method of claim 16, wherein the patient is treated for moderate to severe psoriasis.

29. 17. The method of claim 16, further comprising the step of discontinuing treatment of a patient previously treated with at least one dose of secukinumab and deciding to treat said patient with guselkumab.

30. 30. The method of claim 29, further comprising measuring the psoriasis clinical endpoints PASI90, PASI100, IGA0, and / or IGA1 44 and / or 48 weeks after initial treatment, discontinuing treatment of a patient previously treated with at least one dose of secukinumab, and treating said patient with guselkumab.

31. 17. The method of claim 16, wherein the patient is suffering from psoriatic arthritis.

32. 1. A method of treating moderate to severe plaque psoriasis in an adult patient who is a candidate for systemic treatment or phototherapy, comprising: administering to said patient an antibody against IL-23 in an amount that has been clinically proven safe and has been clinically proven effective; the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 116 and a heavy chain variable region having the amino acid sequence of SEQ ID NO: 106; the dosage is 100 mg administered by subcutaneous injection at weeks 0, 4, and every 8 weeks thereafter; the antibody is at a concentration of 100 mg / mL in a single dose pre-filled syringe; the syringe containing 7.9% (w / v) sucrose, 4.0 mM histidine, 6.9 mM L-histidine monohydrochloride monohydrate, and 0.053% (w / v) polysorbate 80; The diluent is water at normal conditions; said patients are treated with said antibody against IL-23 for at least 44 weeks and show greater efficacy in psoriasis clinical endpoints selected from the group consisting of PASI 90, PASI 100, IGA 0, and IGA 1 than patients treated with the antibody secukinumab. The method.

33. 1. A method of selling and / or promoting an approved pharmaceutical product comprising an antibody against IL-23, comprising: the method comprising advertising a clinical endpoint result at 44 weeks after initial treatment after sequential treatment with the antibody against IL-23 as being superior to a clinical endpoint result at 44 weeks after initial treatment after sequential treatment with secukinumab; The antibody against IL-23 comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 116 and a heavy chain variable region having the amino acid sequence of SEQ ID NO:

106. The method.

Images