Predictors of sustained response after treatment with anti-IL23 specific antibodies

Abstract

Methods for predicting sustained response to IL-23 antibody therapy and treating patients with psoriasis with extended dosing intervals after measuring at least one indicator from the psoriasis patient before and during treatment with an IL-23 antibody are useful for treating psoriasis.

Description

[Technical field]

[0001] (Sequence Listing) This application has been submitted electronically in ASCII format and contains a Sequence Listing, which is incorporated herein by reference in its entirety. The ASCII copy was created on July 16, 2019, has the filename JBI6008WOPCT1Seqlist.txt, and is 4,096 bytes in size.

[0002] FIELD OF THEINVENTION The present invention relates to a method for predicting the maintenance of response to IL-23 antibody therapy and treating patients with psoriasis. In particular, the present invention relates to a method for predicting maintenance therapy with IL-23 antibody after measuring at least one indicator from a psoriasis patient before and during treatment with an IL-23 antibody. [Background technology]

[0003] Interleukin (IL)-12 is a secreted heterodimeric cytokine composed of two disulfide-linked glycosylated protein subunits (designated p35 and p40 for their approximate molecular weight). IL-12 is produced primarily by antigen-presenting cells and promotes cellular immunity by binding to a two-chain receptor complex expressed on the surface of T cells or natural killer (NK) cells. The IL-12 receptor beta-1 (IL-12Rβ1) chain binds to the p40 subunit of IL-12, providing the primary interaction between IL-12 and its receptor. However, it is IL-12p35 ligation of the second receptor chain, IL-12Rβ2, that confers intracellular signaling (e.g., STAT4 phosphorylation) and activation of receptor-bearing cells (Presky et al, 1996). IL-12 signaling in parallel with antigen presentation is thought to trigger T cell differentiation towards the T helper 1 (Th1) phenotype, characterized by interferon gamma (IFNγ) production (Trinchieri, 2003). Th1 cells are thought to promote immunity against several intracellular pathogens, generate complementary antibody isotypes, and contribute to tumor immunosurveillance. This makes IL-12 a critical component of the host defense immune mechanism.

[0004] It was discovered that the p40 protein subunit of IL-12 can also associate with a separate protein subunit designated p19 to form a new cytokine, IL-23 (Oppman et al, 2000). IL-23 also signals through a two-chain receptor complex. The p40 subunit is shared between IL-12 and IL-23, and thus the IL-12Rβ1 chain is also shared between IL-12 and IL-23. However, it is IL-23p19 ligation of the second component of the IL-23 receptor complex, IL-23R, that confers IL-23-specific intracellular signaling (e.g., STAT3 phosphorylation) and subsequent IL-17 production by T cells (Parham et al, 2002; Aggarwal et al. 2003). Recent studies have shown that the biological functions of IL-23 and IL-12 are distinct, despite the structural similarities between these two cytokines (Langrish et al, 2005).

[0005] Abnormal regulation of IL-12 and Th1 cell populations has been linked to many immune-mediated diseases, as neutralization of IL-12 with antibodies has been effective in treating animal models of psoriasis, multiple sclerosis (MS), rheumatoid arthritis, inflammatory bowel disease, insulin-dependent (type 1) diabetes mellitus, and uveitis (Leonard et al., 1995; Hong et al., 1999; Malfait et al., 1998; Davidson et al., 1998). However, these studies targeted the shared p40 subunit, so both IL-12 and IL-23 were neutralized in vivo. Thus, it is unclear whether IL-12 or IL-23 mediated the disease or whether both cytokines need to be inhibited to achieve disease suppression. Recent studies have confirmed that IL-23 inhibition can provide benefits comparable to anti-IL-12p40 strategies, either in IL-23p19-deficient mice or with specific antibody neutralization of IL-23 (Cua et al., 2003; Murphy et al., 2003; Benson et al., 2004). Thus, there is growing evidence for a specific role of IL-23 in immune-mediated diseases. Neutralizing IL-23 without inhibiting the IL-12 pathway could provide an effective therapy for immune-mediated diseases with limited impact on important host defense immune mechanisms. This would represent a significant improvement over current treatment options.

[0006] Psoriasis is a common chronic immune-mediated skin disease with significant comorbidities such as psoriatic arthritis (PsA), depression, cardiovascular disease, hypertension, obesity, diabetes, metabolic syndrome, and Crohn's disease. Plaque psoriasis is the most common form of the disease and presents with sharply demarcated erythematous lesions covered with silvery white scales. The plaques are often pruritic, painful, disfiguring and crippling, and a significant proportion of psoriasis patients have plaques on the hands / nails, face, feet, and genitals. Thus, psoriasis adversely affects health-related quality of life (HRQoL) to a significant extent, including imposing physical and psychosocial burdens that go beyond the physical skin symptoms and interfere with daily activities. For example, psoriasis adversely affects family, spouse, social, and work relationships and is associated with a high incidence of depression and increased suicidal tendencies.

[0007] Histological characterization of psoriatic lesions has revealed epidermal thickening due to abnormal keratinocyte proliferation and differentiation, as well as dermal infiltration and colocalization of CD3+ T lymphocytes and dendritic cells. Although the epidemiology of psoriasis is not well defined, gene and protein analysis has shown that IL-12, IL-23, and their downstream molecules are overexpressed in psoriatic lesions and may in part correlate with the severity of psoriatic disease. Some therapies used to treat psoriasis regulate the levels of IL-12 and IL-23, which are speculated to contribute to their efficacy. Th1 and Th17 cells can produce effector cytokines that induce the production of vasodilators, chemoattractants, and the expression of adhesion molecules on endothelial cells, which in turn promotes the recruitment of monocytes and neutrophils, T cell infiltration, angiogenesis, and keratinocyte activation and hyperplasia. Activated keratinocytes can produce chemoattractant factors that promote neutrophil, monocyte, T cell, and dendritic cell trafficking, thus establishing a cycle of inflammation and keratinocyte hyperproliferation.

[0008] Elucidation of the pathogenesis of psoriasis has led to effective biologic therapies targeting both tumor necrosis factor-alpha (TNF-α), interleukin (IL)-12 and IL-23, and most recently IL-17 and IL-23 alone (including Phase 1, 2 and 3 clinical trials with guselkumab). Guselkumab (also known as CNTO1959) is a fully human IgG1 lambda monoclonal antibody that binds to the p19 subunit of IL-23 and inhibits the intracellular and downstream signaling of IL-23 required for terminal differentiation and maintenance of T helper (Th)17 cells and CD8+IL17A producing cells (Tc17 cells). Summary of the Invention [Means for solving the problem]

[0009] In a first aspect, the present invention relates to a method for predicting sustained response to IL-23 antibody therapy and treating a patient with psoriasis, the method comprising: administering to said patient an IL-23 antibody; b. A biological specimen is or has been obtained from the patient; and c. measuring or having measured at least one indicator from the patient or the biological sample before and during treatment with the IL-23 antibody, the indicator being selected from the group consisting of a clinical parameter, a pharmacokinetic marker, and a serum biomarker; d. generating a predictive patient profile based on the measured indicators; and e. administering said maintenance therapy of said IL-23 antibody according to said predicted patient profile.

[0010] In a preferred embodiment, the anti-IL-23 specific antibody is guselkumab.

[0011] In another aspect, the invention relates to a method for predicting sustained response to IL-23 antibody therapy and treating a patient with psoriasis, the method comprising: a. administering guselkumab at a dose of 100 mg subcutaneously to the patient at an initial dose, 4 weeks after the initial dose, and every 8 weeks after the dose at week 4, wherein the patient is identified as a responder to guselkumab and has a PASI90, PASI100, or IGA score of 0 or 1; b. A biological specimen is or has been obtained from the patient; and c. At least one indicator from the patient or biological sample selected from the group consisting of clinical parameters including disease duration, BMI and / or IGA score, and PASI, pharmacokinetic markers including blood concentration of guselkumab, and serum protein biomarkers including IL17F and MIP1β, is or has been measured at baseline or at 28 weeks of treatment before and during treatment with guselkumab. d. generating a predictive patient profile based on the measured indicators; and e. administering the maintenance therapy of the IL-23 antibody according to the predicted patient profile, wherein the maintenance therapy is administered at a dosing interval selected from the group consisting of: greater than every 8 weeks, every 12 weeks, every 16 weeks, every 20 weeks, every 24 weeks, every 28 weeks, every 32 weeks, every 36 weeks, every 40 weeks, every 44 weeks, and every 48 weeks. [Brief description of the drawings]

[0012] [Figure 1] Design of the 48-week VOYAGE 2 trial and the proportion of patients in arms 1c and 2b of guselkumab-treated PsO patients who achieved long-term maintenance of PASI90 response. [Diagram 2] Shorter disease duration was associated with longer maintenance of PASI90 response: mean disease duration at baseline was 19.4 years in patients without a PASI90 response ("N", n=115) versus 14.5 years in patients with a PASI90 response at week 48 ("Y", n=67), respectively (p<0.005, Welch's t-test). [Diagram 3]Lower BMI at baseline was associated with longer-term maintenance of PASI90 response: mean baseline BMI was 29.9 in patients without a PASI90 response at week 48 ("N", n=115) versus 26.9 in patients with a PASI90 response at week 48 ("Y", n=67), respectively (p<0.001, Welch's t-test). [Figure 4] Achieving IGA0 at week 28 was associated with long-term maintenance of PASI90 response: 45.5% of patients who achieved IGA0 at week 28 had a PASI90 response at week 48 compared with 19.7% of patients who did not achieve IGA0 at week 28 who did not have a PASI90 response at week 48 (odds ratio of 3.38, p<0.001, Fisher's exact test). [Diagram 5] Achieving a PASI100 response at week 28 was associated with prolonged maintenance of a PASI90 response: 45.8% of patients who achieved a PASI100 response at week 28 had a PASI90 response at week 48 compared with 24% of patients who did not achieve a PASI100 response at week 28 (odds ratio of 2.66, p<0.005, Fisher's exact test). [Figure 6] Performance of the model using disease duration as the sole predictor in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.619 and 0.622 for the training set (group 1c) and test set (group 2b), respectively. [Figure 7] Performance of the model using baseline BMI as the sole predictor in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.626 and 0.588 for the training set (group 1c) and test set (group 2b), respectively. [Figure 8]Performance of the model using IGA0 status at week 28 as the single predictor in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.624 and 0.642 for the training set (group 1c) and test set (group 2b), respectively. [Figure 9] Performance of the model using PASI100 response at week 28 as the sole predictor in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.614 and 0.627 for the training set (group 1c) and test set (group 2b), respectively. [Figure 10] Lower serum levels of IL17F at baseline were associated with longer-term maintenance of PASI90 response: the geometric mean serum levels of IL17F at baseline was 6.9 pg / mL in patients without a PASI90 response at week 48 ("N", n=55) versus 4.5 pg / mL in patients with a PASI90 response at week 48 ("Y", n=67), respectively (p<0.03, Welch's t-test). [Figure 11] Lower serum levels of MIP1β at baseline were associated with longer-term maintenance of PASI90 response: the geometric mean serum levels of MIP1β at baseline was 114 pg / mL in patients without a PASI90 response at week 48 ("N", n=53) versus 92 pg / mL in patients with a PASI90 response at week 48 ("Y", n=63), respectively (p<0.02, Welch's t-test). [Figure 12] Performance of single predictor models based on baseline serum levels of IL17F or MIP1β in predicting long-term maintenance of PASI90 response in the training set (Group 1c). Predictive power values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.629 and 0.665, respectively, using baseline IL17F or MIP1β as single predictors. [Figure 13]Higher guselkumab blood concentrations at week 28 were associated with longer-term maintenance of PASI90 response. Mean guselkumab blood concentrations were 0.98 μg / mL in patients without a PASI90 response at week 48 ("N", n=114) versus 1.70 μg / mL in patients with a PASI90 response at week 48 ("Y", n=66), respectively (p<1E-07, Welch's t-test). [Figure 14] Performance of the model using guselkumab blood concentration at week 28 as the sole predictor in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.770 and 0.634 for the training set (group 1c) and test set (group 2b), respectively. [Figure 15] Performance of single predictor models based on disease duration (model 1), baseline body mass index (BMI, model 2), baseline serum levels of IL17F (model 3) or MIP1β (model 4), achievement of Psoriasis Area and Severity Index (PASI) 100 at week 28 (model 5), achievement of Investigator Global Assessment (IGA) clearance at week 28 (model 6), and blood guselkumab (GUS) levels at week 28 (model 7) in predicting long-term maintenance of PASI90 response in the training set (group 1c). The predictive ability of each model is evaluated as the area under the curve (AUC) of the receiver operating characteristic curve. [Figure 16] Performance of predictive models based on multiple baseline predictors, including disease duration, baseline body mass index (BMI), baseline serum levels of IL17F or MIP1β, in predicting long-term maintenance of PASI90 response in the training set (group 1c). The predictive ability of each model is assessed as the area under the curve (AUC) of the receiver operating characteristic curve. [Figure 17]Performance of prediction models based on multiple predictors, including disease duration, baseline body mass index (BMI), baseline serum levels of IL17F or MIP1β, achieving investigator's global assessment (IGA) clearance at week 28, and blood guselkumab (GUS) levels at week 28, in predicting long-term maintenance of PASI90 response in the training set (group 1c). The predictive ability of each model is evaluated as the area under the curve (AUC) of the receiver operating characteristic curve. [Figure 18] Performance of the model combining disease duration, baseline BMI, and IGA0 response at week 28 as predictors in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.724 for the training set (group 1c) and 0.712 for the test set (group 2b), respectively. [Figure 19] Performance of the model combining disease duration, IGA0 response, and blood guselkumab (GUS) levels at week 28 as predictors in predicting long-term maintenance of PASI90 response. The predictive ability values, assessed as the area under the curve (AUC) of the receiver operating characteristic curve, are 0.829 for the training set (group 1c) and 0.738 for the test set (group 2b), respectively. [Figure 20]A logistic regression model based on disease duration (PSODUR), baseline body mass index (BMI), and achieving investigator global assessment (IGA) skin clearance at week 28 (IGA0_WK28) was generated to predict maintenance of a PASI score <3 by administering maintenance guselkumab therapy at 16-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI90 response at week 28. The equation for the logistic regression model is defined as predicted score=1 / (1+exp(-(3.3492-0.03244×PSODUR-0.05807×BMI+1.6254×IGA0_WK28))). The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 67% of guselkumab-treated patients had a predictive score greater than 0.72, and 90% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 16-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28. [Figure 21]A logistic regression model based on disease duration (PSODUR), baseline body mass index (BMI), and achieving investigator's global assessment (IGA) clearance at week 28 (IGA0_WK28) was generated to predict maintenance of a PASI score <3 by administering maintenance guselkumab therapy at 20-week dosing intervals following initial doses of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI90 response at week 28. The equation for the logistic regression model is defined as predicted score=1 / (1+exp(-(2.6101-0.02557×PSODUR-0.05892×BMI+1.0128×IGA0_WK28))). The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 33% of guselkumab-treated patients had a predictive score greater than 0.79, and 85% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 20-week dosing intervals following the initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28. [Figure 22]A logistic regression model based on disease duration (PSODUR), achieving investigator's global assessment (IGA) clearance at week 28 (IGA0_WK28), and blood guselkumab levels at week 28 (GUS_WK28) was generated to predict maintenance of a PASI score <3 by administering maintenance guselkumab therapy at 20-week dosing intervals following initial doses of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI90 response at week 28. The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(-0.1615-0.02004 x PSODUR + 1.0145 x IGA0_WK28 + 0.8199 x GUS_WK28))). The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 42% of guselkumab-treated patients had a predictive score greater than 0.75, and 89% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 20-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28. [Figure 23]A logistic regression model based on disease duration (PSODUR), achieving investigator's global assessment (IGA) clearance at week 28 (IGA0_WK28), and blood guselkumab levels at week 28 (GUS_WK28) was generated to predict maintenance of a PASI score <3 by administering maintenance guselkumab therapy at 24-week dosing intervals following initial doses of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI90 response at week 28. The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(-1.1498-0.02462 x PSODUR + 1.2800 x IGA0_WK28 + 0.9031 x GUS_WK28))). The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 40% of guselkumab-treated patients had a predictive score greater than 0.60, and 78% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 24-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] As used herein, methods of treating psoriasis include the administration of isolated, recombinant and / or synthetic anti-IL-23 specific human antibodies and diagnostic and therapeutic compositions, methods, and devices.

[0014] As used herein, "anti-IL-23 specific antibodies," "anti-IL-23 antibodies," "antibody portions," or "antibody fragments" and / or "antibody variants," and the like, include any protein or peptide-containing molecule comprising at least a portion of an immunoglobulin molecule, such as, but not limited to, at least one complementarity determining region (CDR) or ligand binding portion thereof of a heavy or light chain, a heavy or light chain variable region, a heavy or light chain constant region, a framework region, or any portion thereof, or at least a portion of an IL-23 receptor or binding protein, that can be incorporated into an antibody of the invention. Such antibodies optionally further affect a specific ligand, for example, but not limited to, such antibodies modulate, reduce, enhance, antagonize, promote, reduce, ameliorate, block, inhibit, neutralize, and / or interfere with at least one IL-23 activity or binding, or IL-23 receptor activity or binding, in vitro, in situ, and / or in vivo. As non-limiting examples, suitable anti-IL-23 antibodies, specified portions or variants of the invention can bind to at least one IL-23 molecule or a specified portion, variant or domain thereof. Suitable anti-IL-23 antibodies, specified portions or variants can also optionally affect at least one IL-23 activity or function, such as, but not limited to, RNA, DNA, or protein synthesis, IL-23 release, IL-23 receptor signaling, membrane IL-23 cleavage, IL-23 activity, IL-23 production and / or synthesis.

[0015] The term "antibody" is further intended to encompass antibodies, digested fragments thereof, specified portions, and variants, including antibody mimetics or portions of antibodies that mimic the structure and / or function of antibodies, such as single chain antibodies and fragments thereof. Functional fragments include antigen-binding fragments that bind to mammalian IL-23. For example, antibody fragments capable of binding to IL-23 or portions thereof, including but not limited to Fab (e.g., by papain digestion), Fab' (e.g., by pepsin digestion and partial reduction), and F(ab')2 (e.g., by pepsin digestion), facb (e.g., by plasmin digestion), pFc' (e.g., by pepsin or plasmin digestion), Fd (e.g., by pepsin digestion, partial reduction, and reassembly), Fv, or scFv (e.g., by molecular biology techniques) fragments are encompassed by the present invention (see, e.g., Colligan, Immunology, supra).

[0016] Such fragments can be produced by enzymatic cleavage, synthetic or recombinant techniques, as known in the art and / or as described herein. Antibodies can also be produced in a variety of truncated forms using antibody genes in which one or more stop codons have been introduced upstream of the natural stop site. For example, a combination of genes encoding a F(ab')2 heavy chain portion can be produced by inserting the C(A) codon into the heavy chain. H The antibody can be designed to contain DNA sequences encoding a region of the antibody that binds to the antibody, such as a nucleotide sequence encoding the nucleotide sequence of the antibody, a region encoding the nucleotide sequence of the antibody, and / or a hinge region. The various portions of the antibody can be joined chemically by conventional techniques, or can be prepared as a contiguous protein using genetic engineering techniques.

[0017] As used herein, the term "human antibody" refers to an antibody that is a human antibody that comprises substantially all parts of the protein (e.g., CDRs, frameworks, C L , C H Domain (e.g., C H 1. C H 2. C H 3), Hinge (V L , V H) refers to an antibody that is substantially non-immunogenic in humans with only minor sequence changes or mutations. A "human antibody" may also be an antibody derived from or closely matched to a human germline immunoglobulin sequence. A human antibody may contain amino acid residues not encoded by a germline immunoglobulin sequence (e.g., mutations introduced by random or site-specific mutagenesis in vivo or by somatic mutation in vivo). In many cases, this means that the human antibody is substantially non-immunogenic in humans. Human antibodies have been classified into groups based on the similarity of their amino acid sequences. Thus, using sequence similarity searches, antibodies with similar linear sequences can be selected as templates for creating human antibodies. Similarly, antibodies with names including primates (monkeys, baboons, chimpanzees, etc.), rodents (mouse, rats, rabbits, guinea pigs, hamsters, etc.) and other mammals are indicative of specific antibodies of such species, subgenus, genus, subfamily, and family. Furthermore, chimeric antibodies may include any combination of the above. Such changes or mutations optionally and preferably retain or reduce immunogenicity in humans or other species compared to the unaltered antibody. Thus, a human antibody differs from a chimeric antibody or a humanized antibody.

[0018] It is noted that human antibodies may be produced by non-human animals or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. Furthermore, when a human antibody is a single-chain antibody, it may contain a linker peptide not found in naturally occurring human antibodies. For example, an Fv may contain a linker peptide, such as 2 to about 8 glycine or other amino acid residues, connecting the variable region of the heavy chain and the variable region of the light chain. Such a linker peptide is considered to be of human origin.

[0019] Bispecific, heterospecific, heterobinding or similar antibodies may also be used, which are monoclonal, preferably human or humanized, antibodies with binding specificities for at least two different antigens, one of which is for at least one IL-23 protein and the other for any other antigen. Methods for producing bispecific antibodies are known in the art. Traditionally, recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-light chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537 (1983)). Due to the random combination of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a possible mixture of 10 different antibody molecules, of which only one has the correct bispecific structure. Purification of the correct molecule (usually done by affinity chromatography steps) is quite laborious and the product yield is low. Similar procedures are described, for example, in WO 93 / 08829, U.S. Pat. Nos. 6,210,668, 6,193,967, 6,132,992, 6,106,833, 6,060,285, 6,037,453, 6,010,902, 5,989,530, 5,959,084, 5,959,083 ... ,932,448, 5,833,985, 5,821,333, 5,807,706, 5,643,759, 5,601,819, 5,582,996, 5,496,549, 4,676,980, WO 91 / 00360, WO 92 / 00373, EP 03089, Traunecker et al., EMBO J. 10:3655 (1991), and Suresh et al., Methods in Enzymology 121:210 (1986), each of which is incorporated herein by reference in its entirety.

[0020] Anti-IL-23 specific antibodies (also referred to as IL-23 specific antibodies) (or antibodies against IL-23) useful in the methods and compositions of the invention may optionally be characterized by high affinity binding to IL-23, and optionally and preferably low toxicity. In particular, antibodies of the invention, specified fragments or variants thereof, in which the individual components such as the variable region, constant region, and framework, individually and / or collectively, optionally and preferably, have low immunogenicity, are useful in the present invention. Antibodies that can be used in the present invention may optionally be characterized by the ability to treat patients for extended periods with measurable alleviation of symptoms and low and / or acceptable toxicity. Low or acceptable immunogenicity and / or high affinity, as well as other favorable properties, may contribute to the therapeutic results obtained. "Low immunogenicity" is defined herein as a significant increase in HAHA, HACA or HAMA responses in less than about 75%, or preferably less than about 50%, of treated patients, and / or a low titer (less than about 300, preferably less than about 100, as measured by double antigen enzyme immunoassay) in treated patients (Elliott et al., Lancet 344:1125-1127 (1994), incorporated herein by reference in its entirety). "Low immunogenicity" can also be defined as the incidence of titer levels of antibodies to anti-IL-23 antibodies in patients treated with anti-IL-23 antibodies as occurring in less than 25% of patients treated at the recommended dose, preferably less than 10% of patients treated, during the recommended course of therapy during the treatment period.

[0021] The term "treat" or "treatment" and similar terms herein means to obtain a desired pharmacological, physiological, or therapeutic effect. The effect may be prophylactic in terms of preventing or partially preventing a disease, symptom, or condition, and / or may be therapeutic in terms of partially or completely curing a disease, symptom, condition, or adverse effects resulting from the disease. The term "treatment" as used herein encompasses any treatment of a disease in a mammal, particularly a human, including (a) preventing the disease from occurring in a subject who may be predisposed to the disease but has not yet been diagnosed as having the disease, (b) inhibiting the disease, i.e., arresting its progression, or (c) relieving the disease, i.e., causing the disease and / or its symptoms or conditions to regress. The present invention is directed to the treatment of patients suffering from diseases related to psoriasis. The present invention is concerned with preventing, inhibiting, or relieving adverse effects resulting from psoriasis over a long period of time and / or caused by physiological responses to psoriasis present in a biological system over a long period of time.

[0022] The term "biological sample" encompasses a variety of sample types obtained from an organism and can be used in diagnostic or monitoring assays. The term encompasses blood samples and other liquid samples of biological origin, solid tissue samples such as biopsy specimens or tissue cultures or cells derived therefrom, and their progeny. The term encompasses samples that have been manipulated in any way after obtaining the sample, such as by treatment with reagents, solubilization, or enrichment for specific components. The term encompasses clinical samples, and also includes cells in cell cultures, cell supernatants, cell lysates, serum, plasma, biological fluids, and tissue samples. The term "biological sample" is meant to distinguish samples in a clinical setting from samples that may be recombinant samples or may be derived from recombinant samples.

[0023] As used herein, a "patient" is a human in need of at least one prophylactic or therapeutic agent. In a preferred embodiment, the prophylactic or therapeutic agent comprises an IL-23 antibody.

[0024] In an exemplary embodiment, each patient has a unique patient profile that lists personal information about the patient (such as name, address, etc.) as well as details of the patient's medication information, including the patient's medication schedule. In one embodiment, the drug comprises an IL-23 antibody. As used herein, a patient profile further comprises any measurements collected from the patient before and during administration of an IL-23 antibody. In one embodiment, the collected measurements comprise at least one clinical parameter. In a preferred embodiment, the clinical parameters included in the database of the present invention may include, but are not limited to, psoriasis duration, BMI, IGA score. PASI and any other standard clinical findings are performed by a physician. In another embodiment, the collected measurements comprise at least one pharmacokinetic marker. In a preferred embodiment, the pharmacokinetic marker is the blood concentration of an IL-23 antibody. In another embodiment, the collected measurements comprise at least one serum protein biomarker. In a preferred embodiment, the serum protein biomarkers measured may include, but are not limited to, IL17F and MIP1β.

[0025] In the context of the present invention, a "marker" corresponds to a protein or protein fragment present in a biological sample from a patient, the amount of the marker in the biological sample providing information regarding whether the patient exhibits a change in biological state, such as the presence or absence of a disease or an improvement or reduction in a disease state. In one described embodiment, the marker is a pharmacokinetic marker. In another embodiment, the marker is a serum protein marker.

[0026] usefulness The isolated nucleic acids of the present invention can be used to produce at least one anti-IL-23 antibody or specific variants thereof, which can be used to measure or act on cells, tissues, organs or animals (including mammals and humans) to diagnose, monitor, regulate, treat, alleviate, help prevent the incidence of, or reduce the symptoms of psoriasis.

[0027] Such methods may include administering an effective amount of a composition or pharmaceutical composition comprising at least one anti-IL-23 antibody to a cell, tissue, organ, animal, or patient in need of such modulation, treatment, amelioration, prevention, or reduction of a symptom, effect, or mechanism. An effective amount may include an amount of about 0.001-500 mg / kg per single (e.g., bolus), multiple, or continuous administration, or an amount that achieves a serum concentration of 0.01-5000 μg / mL per single, multiple, or continuous administration, or any effective range or value therein, as determined using known methods, as described herein or known in the relevant art.

[0028] References All publications or patents cited herein, whether specifically stated or not, are incorporated herein by reference in their entirety to indicate the prior art at the time of the invention and / or to describe and enable the invention. A publication refers to any scientific publication or patent publication, or any other information available in any media format, including all recorded in electronic or printed format. The following publications are incorporated by reference in their entirety: Ausubel, et al., eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, NY (1987-2001); Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, NY (1989); Harlow and Lane, antibodies, a Laboratory Manual, Cold Spring Harbor, NY (1989); Colligan, et al., eds., Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001); Colligan et al., Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001).

[0029] Antibodies of the Invention - Production and Purification The at least one anti-IL-23 used in the methods of the present invention can optionally be produced by a cell line, a mixed cell line, an immortalized cell, or a clonal population of immortalized cells, as known in the art. See, e.g., Ausubel, et al., eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, NY (1987-2001); Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, NY (1989); Harlow and Lane, antibodies, a Laboratory Manual, Cold Spring Harbor, NY (1989); Colligan, et al., eds., Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001); Colligan et al., Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001), each of which is incorporated by reference in its entirety.

[0030] A preferred anti-IL-23 antibody is guselkumab (also referred to as CNTO1959), which has a heavy chain variable region amino acid sequence of SEQ ID NO:7, a light chain variable region amino acid sequence of SEQ ID NO:8, and has heavy chain CDR amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and light chain CDR amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6. Other anti-IL-23 antibodies have sequences listed herein and are described in U.S. Patent No. 7,935,344, which is incorporated herein by reference in its entirety.

[0031] Human antibodies specific for human IL-23 protein or fragments thereof can be raised against suitable immunogenic antigens, such as isolated IL-23 protein and / or portions thereof (including synthetic molecules such as synthetic peptides). Other specific or general mammalian antibodies can be raised as well. Preparation of immunogenic antigens and generation of monoclonal antibodies can be carried out using any suitable technique.

[0032] In one approach, a suitable immortal cell line (e.g., but not limited to, Sp2 / 0, Sp2 / 0-AG14, NSO, NS1, NS2, AE-1, L.5, L243, P3X63Ag8.653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 SA5, U937, MLA144, ACT IV, MOLT4, DA-1, JURKAT, WEHI, K-562, COS, RAJI, NIH 3T3, HL-60, MLA144, NAMALWA, NEURO Myeloma cell lines such as 2A, or heteromyelomas, fusion products thereof, or any cells or fusion cells derived therefrom, or any other suitable cell line known in the art) (see, e.g., www.atcc.org, www.lifetech.com, etc.), antibody producing cells such as, but not limited to, isolated or cloned spleen, peripheral blood, lymph, tonsil, or other immune or B cell containing cells, or recombinant or endogenous, viral, bacterial, algal, progenitor, or progenitor cells, as endogenous or heterologous nucleic acids. The hybridomas are produced by fusing with any other cell expressing the heavy or light chain constant or variable, or framework or CDR sequences, either as a prokaryote, amphibian, insect, reptile, fish, mammal, rodent, horse, ovine, caprine, ovine, primate, eukaryote, genomic DNA, cDNA, rDNA, mitochondrial DNA or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single, double or triple stranded, hybridized, etc., or as any combination thereof. See, e.g., Ausubel and Colligan, Immunology chapter 2, supra, incorporated herein by reference in its entirety.

[0033] Antibody producing cells can also be obtained from peripheral blood, or preferably from the spleen or lymph nodes, of humans or other suitable animals immunized with the antigen of interest. Any other suitable host cells can also be used to express heterologous or endogenous nucleic acid encoding the antibodies, specified fragments or variants thereof of the invention. Fused cells (hybridomas) or recombinant cells can be isolated using selective culture conditions or other suitable known methods and cloned by limiting dilution or cell sorting or other known methods. Cells producing antibodies with the desired specificity can be selected by a suitable assay (e.g., ELISA).

[0034] 2. Select recombinant antibodies from peptide or protein libraries (e.g., but not limited to, bacteriophage, ribosomal, oligonucleotide, RNA, cDNA, and other display libraries, e.g., Cambridge Antibody Technologies, Cambridgeshire, UK; MorphoSys, Martinsreid / Planegg, DE; Biovation, Aberdeen, Scotland, UK; BioInvent, Lund, Sweden; Dyax Corp., Enzon, Affymax / Biosite, Xoma, Berkeley, CA, Ixsys. For example, European Patent No. 368,684, International Application No. PCT / GB91 / 01134, International Application No. PCT / GB92 / 01755, International Application No. PCT / GB92 / 002240, International Application No. PCT / GB92 / 00883, International Application No. PCT / GB93 / 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 PCT / GB97 / 01835, (CAT / MRC), International Publication No. WO 90 / 14443, International Publication No. WO 90 / 14424, International Publication No. WO 90 / 14430, International Application PCT / US94 / 1234, International Publication No. WO 92 / 18619, International Publication No. WO 96 / 07754, (Scripps), International Publication No. WO 96 / 13583, International Publication No. WO 97 / 08320 (MorphoSys), WO 95 / 16027 (BioInvent), WO 88 / 06630, WO 90 / 3809 (Dyax), U.S. Patent No. 4,704,692 (Enzon), International Application PCT / US91 / 02989 (Affymax), WO 89 / 06283, European Patent No. 371998, European Patent No. 550400, (Xoma), European No. 229046, International Application PCT / US91 / 07149 (Ixsys), or stochastically generated peptides or proteins - U.S. Pat. Nos. 5,723,323, 5,763,192, 5,814,476, 5,817,483, 5,824,514, 5,976,862, WO 86 / 05803, EP 590689 (Ixsys, Applied Molecular Evolution (AME) predecessors, each of which is incorporated herein by reference in its entirety), or rely on immunization of transgenic animals capable of producing a repertoire of human antibodies as known in the art and / or described herein (e.g., SCID mice, Nguyen et al., Microbiol. Immunol. 41:901-907 (1997); Samdhu et al., Crit. Rev. Biotechnol. 16:95-118 (1996); Eren et al.Other suitable methods for producing or isolating antibodies of the required specificity can be used, including, but not limited to, the methods described in Immunol. 93:154-161 (1998) (each of which is incorporated by reference in its entirety), and related patents and applications. Such techniques include ribosome display (Hanes et al., Proc. Natl. Acad. Sci. USA, 94:4937-4942 (May 1997); Hanes et al., Proc. Natl. Acad. Sci. USA, 95:14130-14135 (Nov. 1998)), single cell antibody generation techniques (e.g., the selected lymphocyte antibody method ("SLAM") (U.S. Pat. No. 5,627,052; Wen et al., J. Immunol. 17:887-892 (1987); Babcook et al., Proc. Natl. Acad. Sci. USA 93:7843-7848 (1996)), gel microdroplets, and flow cytometry (Powell et al., J. Immunol. 1999, 11:111-111 (1998)). al., Biotechnol. 8:333-337(1990); One Cell Systems, Cambridge, MA; Gray et al., J. Imm. Meth. 182:155-163(1995); Kenny et al., Bio / Technol. 13:787-790(1995)), B cell selection (Steenbakkers et al., Molec. Biol. Reports 19:125-134(1994); Jonak et al., Progress Biotech, Vol. 5, In Vitro Immunization in Hybridoma Technology, Borrebaeck, ed., Elsevier Science Publishers BV, Amsterdam, Netherlands(1988)).

[0035] Methods for engineering or humanizing non-human or human antibodies can be used as well and are well known in the art. Generally, a humanized or modified antibody has one or more amino acid residues that are non-human, such as, but not limited to, mouse, rat, rabbit, non-human primate, or other mammal. These non-human amino acid residues are often replaced by residues referred to as "import" residues. Such "import" residues are typically taken from an "import" variable, constant, or other domain of a known human sequence.

[0036] Known human Ig sequences can be found, for example, at www.ncbi.nlm.nih.gov / entrez / query.fcgi, www.ncbi.nih.gov / igblast, www.atcc.org / phage / hdb.html, www.mrc-cpe.cam.ac.uk / ALIGNMENTS.php, www.kabatdatabase.com / top.html, ftp.ncbi.nih.gov / repository / kabat, www.sciquest.com, each of which is incorporated herein by reference in its entirety. / , www.abcam.com / , www.antibodyresource.com / onlinecomp.html, www.public.iastate.edu / ~pedro / research_tools.html, www.whfreeman.com / immunology / CH05 / kuby05.htm, www.hhmi.org / grants / lectures / 1996 / vlab / , www.path.cam.ac.uk / ~mrc7 / mikeimages.html, mcb.harvard.edu / BioLinks / Immunology.html, www.immunologylink.com, pathbox.wustl.edu / ~hcenter / index.html, www.appliedbiosystems.com, www.nal.usda.gov / awic / pubs / antibody, www.m.ehime-u.ac.jp / ~yasuhito / Elisa.html, www.biodesign.com, www.cancerresearchuk.org, www.biotech.ufl.ed u, www.isac-net.org, baserv.uci.kun.nl / ~jraats / links1.html;www.recab.uni-hd.de / immuno.bme.nwu.edu, www.mrc-cpe.cam.ac.uk, www.ibt .unam.mx / vir / V_mice.html, http: / / www.bioinf.org.uk / abs, antibody.bath.ac.uk, www.unizh.ch, www.cryst.bbk.ac.uk / ~ubcg07s, www.nimr.mrc.ac.uk / CC / ccaewg / ccaewg.html, www.path.cam.ac.uk / ~mrc7 / humanisation / TAHHP.html, www.ibt.unam.mx / vir / structure / stat_aim.html, www.biosci.missouri.edu / smithgp / index.html, www.jerini.de, Kabat et al., Sequences of Proteins of Immunological Interest, USDept. Health (1983), each of which is incorporated herein by reference in its entirety.

[0037] Such imported sequences can be used to reduce immunogenicity or to reduce, enhance or modify binding, affinity, association rate constant, dissociation rate constant, avidity, specificity, half-life, or any other suitable property as known in the art. Generally, CDR residues directly and most substantially affect antigen binding. Thus, non-human sequences of variable and constant regions can be replaced with human amino acids or other amino acids while maintaining some or all of the non-human or human CDR sequences.

[0038] Antibodies may optionally be humanized, or human antibodies may be engineered while retaining high affinity for the antigen and other favorable biological properties. To this end, humanized (or human) antibodies may optionally be prepared by a process of analyzing the parental sequences and various theoretical humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformations of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., analysis of residues that influence the antigen-binding ability of the candidate immunoglobulin. In this way, framework (FR) residues can be selected and combined from the consensus and import sequences such that desired antibody properties, such as enhanced affinity for the target antigen(s), are achieved.

[0039] In addition, a human IL-23 specific antibody used in the methods of the invention may comprise a human germline light chain framework. In certain embodiments, the light chain germline sequence is selected from a human VK sequence, including, but not limited to, A1, A10, A11, A14, A17, A18, A19, A2, A20, A23, A26, A27, A3, A30, A5, A7, B2, B3, L1, L10, L11, L12, L14, L15, L16, L18, L19, L2, L20, L22, L23, L24, L25, L4 / 18a, L5, L6, L8, L9, O1, O11, O12, O14, O18, O2, O4, and O8. In certain embodiments, the light chain human germline framework is selected from V1-11, V1-13, V1-16, V1-17, V1-18, V1-19, V1-2, V1-20, V1-22, V1-3, V1-4, V1-5, V1-7, V1-9, V2-1, V2-11, V2-13, V2-14, V2-15, V2-17, V2-19, V2-6, V2-7, V2-8, V3-2, V3-3, V3-4, V4-1, V4-2, V4-3, V4-4, V4-6, V5-1, V5-2, V5-4, and V5-6.

[0040] In other embodiments, the human IL-23 specific antibody used in the methods of the invention may comprise a human germline heavy chain framework. In certain embodiments, the heavy chain human germline framework is selected from the group consisting of VH1-18, VH1-2, VH1-24, VH1-3, VH1-45, VH1-46, VH1-58, VH1-69, VH1-8, VH2-26, VH2-5, VH2-70, VH3-11, VH3-13, VH3-15, VH3-16, VH3-20, VH3-21, VH3-23, VH3-30, VH3- 33, VH3-35, VH3-38, VH3-43, VH3-48, VH3-49, VH3-53, VH3-64, VH3-66, VH3-7, VH3-72, VH3-73, VH3-74, VH3-9, VH4-28, VH4-31, VH4-34, VH4-39, VH4-4, VH4-59, VH4-61, VH5-51, VH6-1, and VH7-81.

[0041] 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 subregions, such as FR2 and FR3). In certain embodiments, at least FRL1, FRL2, FRL3, or FRL4 is fully human. In other embodiments, at least FRH1, FRH2, FRH3, or FRH4 is fully human. In some embodiments, at least FRL1, FRL2, FRL3, or FRL4 is a germline sequence (e.g., human germline) or comprises a human consensus sequence for a particular framework (which are readily available at the sources of known human Ig sequences mentioned above). In other embodiments, at least FRH1, FRH2, FRH3, or FRH4 is a germline sequence (e.g., human germline) or comprises a human consensus sequence for a particular framework. In preferred embodiments, the framework regions are fully human framework regions.

[0042] Humanization or engineering of the antibodies of the present invention has been described by Winter (Jones et al., Nature 321:522 (1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al., Science 239:1534 (1988)), Sims et al., J. Immunol. 151:2296 (1993), Chothia and Lesk, J. Mol. Biol. 196:901 (1987), Carter et al., Proc. Natl. Acad. Sci. USA 89:4285 (1992), Presta et al. al., J.Immunol.151-2623 (1993), U.S. Patent No. 5723323, U.S. Patent No. 5976862, U.S. Patent No. 5824514, U.S. Patent No. 5817483, U.S. Patent No. 5814476, U.S. Patent No. 5763192, U.S. Patent No. 5723323, U.S. Patent No. 5 , 766886, 5714352, 6204023, 6180370, 5693762, 5530101, 5585089, 5225539, 4816567, International Application PCT / :US98 / 1 This can be done using any known method, such as, but not limited to, those described in US96 / 18978, US91 / 09630, US91 / 05939, US94 / 01234, GB89 / 01334, GB91 / 01134, GB92 / 01755, WO 90 / 14443, WO 90 / 14424, WO 90 / 14430, EP 229246 (each of which is incorporated by reference in its entirety, including the references cited therein).

[0043] In certain embodiments, the antibody comprises an altered (e.g., mutated) Fc region. For example, in some embodiments, the Fc region is altered to reduce or enhance the effector function of the antibody. In some embodiments, the Fc region is an isotype selected from IgM, IgA, IgG, IgE, or other isotypes. Alternatively, or in addition, it may be useful to combine the amino acid modification with one or more further amino acid modifications that alter the C1q binding and / or complement dependent cytotoxicity function of the Fc region of the IL-23 binding molecule. A starting polypeptide of particular interest may be one that binds C1q and exhibits complement dependent cytotoxicity (CDC). A polypeptide that has existing C1q binding activity, and optionally also has the ability to mediate CDC, may be modified to enhance one or both of these activities. Amino acid modifications that alter C1q and / or modify its complement dependent cytotoxicity function are described, for example, in WO 0042072, incorporated by reference.

[0044] As disclosed above, the Fc region of the human IL-23 specific antibody of the invention can be engineered with altered effector functions, for example, by modifying C1q binding and / or FcγR binding, thereby altering complement-dependent cytotoxicity (CDC) and / or antibody-dependent cell-mediated cytotoxicity (ADCC) activity. An "effector function" serves to activate or reduce a biological activity (e.g., in a subject). Examples of effector functions include, but are not limited to, C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, downregulation of cell surface receptors (e.g., B cell receptor, BCR), and the like. Such effector functions may require the Fc region to bind to a binding domain (e.g., an antibody variable domain) and can be assessed using a variety of test methods (e.g., Fc binding assays, ADCC assays, CDC assays, and the like).

[0045] For example, variant Fc regions of human IL-23 (or anti-IL-23) antibodies can be generated that have improved C1q binding and improved FcγRIII binding (e.g., that have both improved ADCC activity and improved CDC activity). Alternatively, where it is desired to reduce or eliminate effector function, Fc region variants can be engineered to have reduced CDC activity and / or reduced ADCC activity. In other embodiments, only one of these activities may be enhanced, optionally with the other activity reduced at the same time (e.g., to generate Fc region variants with improved ADCC activity and reduced CDC activity, and vice versa).

[0046] Fc mutations can also be engineered and introduced to alter the interaction with the fetal Fc receptor (FcRn) and improve their pharmacokinetic properties. A collection of human Fc variants with improved binding to FcRn has been described (Shields et al., (2001). High resolution mapping of the binding site on human IgG1 for FcγRI, FcγRII, FcγRIII, and FcRn and design of IgG1 variants with improved binding to the FcγR, J. Biol. Chem. 276:6591-6604).

[0047] Another type of amino acid substitution serves to alter the glycosylation pattern of the Fc region of a human IL-23 specific antibody. Glycosylation of the Fc region is typically either N-linked or O-linked. N-linked refers to the addition of a carbohydrate moiety to the side chain of an asparagine residue. O-linked glycosylation refers to the attachment of one of the sugars, N-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used. The recognition sequences for enzymatic addition of a carbohydrate moiety to the asparagine side chain peptide sequence are asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline. Thus, the presence of either of these peptide sequences in a polypeptide provides a site at which glycosylation can occur.

[0048] The glycosylation pattern can be altered, for example, by deleting one or more glycosylation site(s) found in the polypeptide and / or adding one or more glycosylation sites not present in the polypeptide. Addition of glycosylation sites to the Fc region of a human IL-23-specific antibody is conveniently accomplished by modifying the amino acid sequence to include one or more of the above tripeptide sequences (for N-linked glycosylation sites). An exemplary glycosylation variant has an amino acid substitution of residue Asn297 of the heavy chain. This change may also be made by the addition of, or substitution by, one or more serine or threonine residues to the original polypeptide sequence (for O-linked glycosylation sites). Additionally, changing Asn 297 to Ala can remove one of the glycosylation sites.

[0049] In certain embodiments, the human IL-23 specific antibodies of the invention are expressed in cells expressing beta(1,4)-N-acetylglucosaminyltransferase III (GnT III), such that GnT III adds G1cNAc to the human IL-23 antibody. Methods for producing antibodies in such a manner are provided in WO 9954342, WO 03011878, JP 20030003097A1, and Umana et al., Nature Biotechnology, 17:176-180, Feb. 1999, all of which are specifically incorporated herein by reference in their entirety.

[0050] Anti-IL-23 antibodies may also, optionally, be generated by immunization of transgenic animals (e.g., mice, rats, hamsters, non-human primates, etc.) capable of producing a repertoire of human antibodies, as described herein and / or known in the art. Cells producing human anti-IL-23 antibodies may be isolated and immortalized from such animals using suitable methods, such as those described herein.

[0051] Transgenic mice capable of producing a repertoire of human antibodies that bind human antigens can be produced by known methods (see, for example, but not limited to, U.S. Pat. Nos. 5,770,428, 5,569,825, 5,545,806, 5,625,126, 5,625,825, 5,633,425, 5,661,016, and 5,789,650 issued to Lonberg et al.; WO 98 / 50433 to Jakobovits et al.; WO 98 / 24893 to Jakobovits et al.; WO 98 / 24884 to Lonberg et al.; See, for example, WO 97 / 13852 to Lonberg et al., WO 94 / 25585 to Lonberg et al., WO 96 / 34096 to Kucherlapate et al., EP 0463151(B1) to Kucherlapate et al., EP 0710719(A1) to Kucherlapate et al., U.S. Pat. No. 5,545,807 to Surani et al., WO 90 / 04036 to Bruggemann et al., EP 0438474(B1) to Bruggemann et al., EP 0814259(A2) to Lonberg et al., UK Patent 2272440(A) to Lonberg et al., et al.Nature 368:856-859(1994), Taylor et al.,Int.Immunol.6(4)579-591(1994), Green et al.,Nature Genetics 7:13-21(1994), Mendez et al.,Nature Genetics 15:146-156(1997), Taylor et al. al., Nucleic Acids Research 20(23):6287-6295(1992), Tuaillon et al., Proc Natl Acad Sci USA 90(8)3720-3724(1993), Lonberg et al., Int Rev Immunol 13(1):65-93(1995), and Fishwald et al., Nat. Biotechnol 14(7):845-851 (1996), each of which is incorporated herein by reference in its entirety.Generally, these mice contain at least one transgene that comprises DNA derived from at least one human immunoglobulin locus that has been functionally rearranged, or is capable of undergoing functional rearrangement. The endogenous immunoglobulin loci of such mice can be disrupted or deleted to eliminate the ability of the mice to produce antibodies encoded by endogenous genes.

[0052] Screening of antibodies for specific binding to similar proteins or fragments can be successfully accomplished using peptide display libraries. This method involves screening a large collection of peptides for individual members with the desired function or structure. Antibody screening of peptide display libraries is well known in the art. The displayed peptide sequences can be 3-5000 or more amino acids in length, frequently 5-100 amino acids long, and often about 8-25 amino acids long. In addition to direct chemical synthesis methods for generating peptide libraries, several recombinant DNA methods have also been described. One type involves the display of peptide sequences on the surface of bacteriophages or cells. Each bacteriophage or cell contains a nucleotide sequence that codes for a particular displayed peptide sequence. Such methods are described in WO 91 / 17271, WO 91 / 18980, WO 91 / 19818, and WO 93 / 08278.

[0053] Other systems for generating peptide libraries include aspects of both in vitro chemical synthesis and recombinant methods. See WO 92 / 05258, WO 92 / 14843, and WO 96 / 19256. See also U.S. Patent Nos. 5,658,754 and 5,643,768. Peptide display libraries, vectors, and screening kits are commercially available from sources such as Invitrogen (Carlsbad, CA) and Cambridge antibody Technologies (Cambridgeshire, UK). See, e.g., U.S. Patent Nos. 4,704,692, 4,939,666, 4,946,778, 5,260,203, 5,455,030, 5,518,889, 5,534,621, 5,656,730, 5,763,733, 5,767,260, and 5,856,456, all assigned to Enzon; ​​U.S. Patent Nos. 5,223,409, 5,403,484, 5,571,698, and 5,837,500, all assigned to Dyax; U.S. Patent Nos. 5,427,908 and 5,580,717, all assigned to Affymax; Cambridge antibody See US Patent No. 5,885,793 assigned to Genentech, US Patent No. 5,750,373 assigned to Genentech, US Patent No. 5,618,920 assigned to Xoma, US Patent No. 5,595,898, US Patent No. 5,576,195, US Patent No. 5,698,435, US Patent No. 5,693,493, US Patent No. 5,698,417 assigned to Xoma, Colligan, supra, Ausubel, supra, or Sambrook, supra. Each of the above patents and publications is incorporated herein by reference in its entirety.

[0054] The antibodies used in the methods of the present invention can also be prepared using at least one anti-IL-23 antibody encoding nucleic acid to provide a transgenic animal or mammal, such as a goat, cow, horse, sheep, rabbit, etc., that produces such antibodies in its milk. Such animals can be prepared using known methods. See, for example, but not limited to, U.S. Patent Nos. 5,827,690, 5,849,992, 4,873,316, 5,849,992, 5,994,616, 5,565,362, 5,304,489, etc., each of which is incorporated herein by reference in its entirety.

[0055] The antibodies used in the methods of the invention can further be prepared using at least one anti-IL-23 antibody-encoding nucleic acid to provide transgenic plants and cultured plant cells (e.g., but not limited to, tobacco and corn) that produce such antibodies, specified portions, or variants in plant parts or cells cultured therefrom. As a non-limiting example, transgenic tobacco leaves expressing recombinant proteins using, for example, an inducible promoter have been successfully used to provide large quantities of recombinant proteins. See, e.g., Cramer et al., Curr. Top. Microbol. Immunol. 240:95-118 (1999) and references cited therein. Transgenic corn has also been used to express mammalian proteins at commercial production levels with biological activity equivalent to proteins produced in other recombinant systems or purified from natural sources. See, e.g., Hood et al., Adv. Exp. Med. Biol. 464:127-147 (1999) and references cited therein. Antibodies have also been produced in large quantities from transgenic plant seeds, such as tobacco seeds and potato tubers, containing antibody fragments, such as single chain antibodies (scFv). See, e.g., Conrad et al., Plant Mol. Biol. 38:101-109 (1998) and references cited therein. Thus, the antibodies of the present invention can also be produced using transgenic plants according to known methods. See, e.g., Fischer et al., Biotechnol. Appl. Biochem. 30:99-108 (Oct., 1999), Ma et al., Trends Biotechnol. 13:522-7 (1995), Ma et al., Plant Physiol. 109:341-6 (1995), Whitelam et al., Biochem. Soc. Trans. 22:940-944 (1994) and references cited therein. Each of the above documents is incorporated herein by reference in its entirety.

[0056] The antibodies used in the methods of the present invention have a wide range of affinities (K D ) can bind human IL-23. In a preferred embodiment, the human mAb can optionally bind human IL-23 with high affinity. For example, the human mAb can bind human IL-23 with approximately 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 therein, including, but not limited to, K D can be combined with

[0057] The affinity or avidity of an antibody for an antigen can be determined experimentally using any suitable method. (See, e.g., Berzofsky, et al., "Antibody-Antigen Interactions," In Fundamental Immunology, Paul, WE, Ed., Raven Press: New York, NY (1984); Kuby, Janis Immunology, WH Freeman and Company: New York, NY (1992), and methods described therein.) The affinity measured for a particular antibody-antigen interaction may differ when measured under different conditions (e.g., salt concentration, pH). Thus, affinity and other antigen-binding parameters (e.g., K D、 K a , K d Measurements of ) are preferably made using standard solutions of antibody and antigen, and standard buffers, such as those described herein.

[0058] nucleic acid molecule Using the information provided herein, such as, for example, a nucleotide sequence encoding at least 70-100% of the contiguous amino acids of at least one of the light or heavy chain variable or CDR regions described herein, an identified fragment, variant, or consensus sequence thereof, or a deposited vector containing at least one of these sequences, among other sequences disclosed herein, a nucleic acid molecule of the invention encoding at least one anti-IL-23 antibody can be obtained using methods described herein or known in the art.

[0059] The nucleic acid molecules of the present invention may be in the form of RNA, such as mRNA, hnRNA, tRNA or any other form, or in the form of DNA, including but not limited to cDNA and genomic DNA obtained by cloning or produced synthetically, or any combination thereof. The DNA may be triple-stranded, double-stranded or single-stranded, or any combination thereof. Any portion of at least one strand of the DNA or RNA may be the coding strand, also known as the sense strand, or the non-coding strand, called the antisense strand.

[0060] The isolated nucleic acid molecules used in the methods of the present invention may include an open reading frame (ORF), optionally having one or more introns, for example, a nucleic acid molecule comprising at least one specified portion of at least one CDR, such as, but not limited to, CDR1, CDR2, and / or CDR3 of at least one heavy or light chain, a nucleic acid molecule comprising a coding sequence of an anti-IL-23 antibody or a variable region, and a nucleic acid molecule comprising a nucleotide sequence that is substantially different from the above-mentioned nucleic acid molecules, but still encodes at least one anti-IL-23 antibody described herein and / or known in the art due to the degeneracy of the genetic code. Of course, the genetic code is well known in the art. Thus, it would be routine for a person skilled in the art to generate such degenerate nucleic acid variants that code for a specific anti-IL-23 antibody used in the methods of the present invention. See, for example, Ausubel et al., supra, and such nucleic acid variants are included in the present invention. Non-limiting examples of isolated nucleic acid molecules include nucleic acids encoding HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3, respectively.

[0061] As described herein, nucleic acid molecules comprising nucleic acids encoding anti-IL-23 antibodies can include, but are not limited to, those that themselves encode the amino acid sequence of an antibody fragment, sequences encoding the full length of an antibody or a portion of an antibody, coding sequences for an antibody, fragment or portion, and additional sequences, such as at least one intron, with or without the aforementioned additional coding sequences, including, but not limited to, non-coding 5' and 3' sequences, e.g., transcribed non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals (e.g., ribosome binding and stability of mRNA), as well as coding sequences for at least one signal leader or fusion peptide, additional coding sequences encoding additional amino acids, e.g., amino acids that provide additional functions. Thus, the antibody coding sequence can be fused to a marker sequence, e.g., a sequence encoding a peptide that facilitates purification of the antibody containing the antibody fragment or portion to which it is fused.

[0062] Polynucleotides that selectively hybridize to the polynucleotides described herein The method of the present invention uses isolated nucleic acids that hybridize under selective hybridization conditions to the polynucleotides disclosed herein. Thus, the polynucleotides of the present embodiment can be used to isolate, detect, and / or quantify nucleic acids that contain such polynucleotides. For example, the polynucleotides of the present invention can be used to identify, isolate, or amplify partial or full-length clones in a deposited library. In some embodiments, the polynucleotides are genomic or cDNA sequences that are isolated or otherwise complementary to cDNAs in a human or mammalian nucleic acid library.

[0063] Preferably, the cDNA library contains at least 80% of full-length sequences, preferably at least 85% or 90% of full-length sequences, more preferably at least 95% of full-length sequences. The cDNA library can be normalized to increase the representation of rare sequences. Low or medium stringency hybridization conditions are typical, but not limited to, using sequences with low sequence identity to the complementary sequence. Optionally, medium and high stringency conditions can be used for sequences with higher identity. Low stringency conditions allow selective hybridization of sequences with about 70% sequence identity and can be used to identify orthologous or paralogous sequences.

[0064] Optionally, the polynucleotide encodes at least a portion of an antibody. The polynucleotide comprises a nucleic acid sequence that is available for selective hybridization to a polynucleotide encoding an antibody of the invention. See, e.g., Ausubel, supra; Colligan, supra, each of which is incorporated herein by reference in its entirety.

[0065] Nucleic acid construction An isolated nucleic acid can be produced using (a) recombinant methods, (b) synthetic techniques, (c) purification techniques, and / or combinations thereof, as are well known in the art.

[0066] The nucleic acid can conveniently contain sequences in addition to the polynucleotide of the invention. For example, a multiple cloning site containing one or more endonuclease restriction sites can be inserted into the nucleic acid to aid in the isolation of the polynucleotide. Also, a translatable sequence can be inserted to aid in the isolation of the translated polynucleotide of the invention. For example, a hexahistidine marker sequence provides a convenient means for purifying the protein of the invention. The nucleic acid of the invention (excluding the coding sequence) is optionally a vector, adapter, or linker for cloning and / or expression of the polynucleotide of the invention.

[0067] Additional sequences can be added to such cloning and / or expression sequences to optimize their function in cloning and / or expression, to aid in the isolation of polynucleotides, or to improve the introduction of polynucleotides into cells. The use of cloning vectors, expression vectors, adapters, and linkers is well known in the art. (See, e.g., Ausubel, supra, or Sambrook, supra.)

[0068] Recombinant methods for constructing nucleic acids Isolated nucleic acid compositions, such as RNA, cDNA, genomic DNA, or any combination thereof, can be obtained from biological sources using any number of cloning methods known to those of skill in the art. In some embodiments, oligonucleotide probes that selectively hybridize under stringent conditions to the polynucleotides of the present invention are used to identify desired sequences in a cDNA or genomic DNA library. Isolation of RNA and construction of cDNA and genomic libraries are well known to those of skill in the art. (See, e.g., Ausubel, supra, or Sambrook, supra.)

[0069] Nucleic Acid Screening and Isolation Methods Probes based on the sequences of the polynucleotides used in the methods of the invention, such as those disclosed herein, can be used to screen cDNA or genomic libraries. Probes can be used to hybridize to genomic DNA or cDNA sequences to isolate homologous genes in the same or different organisms. Those skilled in the art will appreciate that various degrees of hybridization stringency can be used in the assay, and that either the hybridization or the wash medium can be made more stringent. The more stringent the hybridization conditions, the greater the degree of complementarity between the probe and the target at which duplex formation occurs. The degree of stringency can be controlled by one or more of temperature, ionic strength, pH, and the presence of a partially denaturing solvent such as formamide. For example, the stringency of hybridization is successfully altered by changing the polarity of the reaction solution, for example, by manipulating the formamide concentration in the range of 0% to 50%. The degree of complementarity (sequence identity) required for detectable binding varies depending on the stringency of the hybridization medium and / or the wash medium. The degree of complementarity is optimally 100%, or 70-100%, or any range or value therein, however, it should be understood that minor differences in sequence in the probe and primers can be compensated for by reducing the stringency of the hybridization and / or wash medium.

[0070] Methods for amplifying RNA or DNA are well known in the art and can be used in accordance with the present invention without undue experimentation, based on the teachings and guidance provided herein.

[0071] Known methods of DNA or RNA amplification include the polymerase chain reaction (PCR) and related amplification processes (e.g., U.S. Pat. Nos. 4,683,195, 4,683,202, 4,800,159, and 4,965,188 to Mullis et al., 4,795,699 and 4,921,794 to Tabor et al., 5,142,033 to Innis, 5,122,464 to Wilson et al., 5,091,310 to Innis, 5,066,584 to Gyllensten et al., 5,122,592 to Innis, 5,091,310 to Gyllensten et al., 5,066,584 to Gelfand et al., and the like). No. 4,889,818 to Silver et al., No. 4,994,370 to Biswas, No. 4,766,067 to Ringold, No. 4,656,134 to Ringold), and RNA-mediated amplification (U.S. Pat. No. 5,130,238 to Malek et al., under the trade name NASBA) using antisense RNA to a target sequence as a template for double-stranded DNA synthesis, the entire contents of which are incorporated herein by reference. (See, e.g., Ausubel, supra, or Sambrook, supra.)

[0072] For example, the polymerase chain reaction (PCR) technique can be used to amplify the sequences of polynucleotides and related genes used in the methods of the present invention directly from genomic DNA or cDNA libraries. PCR and other in vitro amplification methods can also be useful, for example, for cloning nucleic acid sequences encoding proteins to be expressed, for generating nucleic acids to be used as probes for detecting the presence of desired mRNA in a sample, for sequencing nucleic acids, or for other purposes. Examples of techniques sufficient to guide the skilled artisan through in vitro amplification methods can be found in Berger, supra, Sambrook, supra, and Ausubel, supra, as well as Mullis et al., U.S. Patent No. 4,683,202 (1987), and Innis, et al., PCR Protocols A Guide to Methods and Applications, Eds., Academic Press Inc, San Diego, CA (1990). Commercial kits for genomic PCR amplification are known in the art. See, for example, Advantage-GC Genomic PCR Kit (Clontech). In addition, for example, the T4 gene 32 protein (Boehringer Mannheim) can be used to improve yields of long PCR products.

[0073] Synthetic methods for constructing nucleic acids The isolated nucleic acid used in the method of the present invention can also be prepared by direct chemical synthesis by known methods (see, for example, Ausubel et al., supra). Chemical synthesis generally produces a single-stranded oligonucleotide that can be converted into double-stranded DNA by hybridization with a complementary sequence or by polymerization with a DNA polymerase using the single strand as a template. Those skilled in the art will recognize that although chemical synthesis of DNA can be limited to sequences of about 100 or more bases, longer sequences can be obtained by ligation of shorter sequences.

[0074] Recombinant Expression Cassettes The present invention uses recombinant expression cassettes that contain nucleic acids. A nucleic acid sequence, e.g., a cDNA or genomic sequence encoding an antibody used in the methods of the present invention, can be used to construct a recombinant expression cassette that can be introduced into at least one desired host cell. A recombinant expression cassette typically contains a polynucleotide that is operably linked to a transcription initiation regulatory sequence that directs transcription of the polynucleotide in the intended host cell. Both heterologous and non-heterologous (i.e., endogenous) promoters can be utilized to direct the expression of the nucleic acid.

[0075] In some embodiments, isolated nucleic acids that function as promoters, enhancers, or other elements can be introduced into the appropriate location (upstream, downstream, or within an intron) of a non-heterologous form of a polynucleotide of the invention to up- or down-regulate expression of the polynucleotide. For example, endogenous promoters can be altered by mutation, deletion, and / or substitution in vivo or in vitro.

[0076] Vectors and host cells The present invention also relates to vectors containing the isolated nucleic acid molecules, host cells engineered with the recombinant vectors, and the production of at least one anti-IL-23 antibody by recombinant techniques well known in the art (see, e.g., Sambrook et al., supra; Ausubel et al., supra, each of which is incorporated herein by reference in its entirety).

[0077] Polynucleotide can be optionally linked to a vector that contains a selection marker for propagation in a host.Generally, plasmid vector is introduced into a precipitate such as calcium phosphate precipitate or into a complex with charged lipid.If vector is a virus, it can be packaged in vitro using a suitable packaging cell line and then transduced into host cell.

[0078] The DNA insert should be operably linked to a suitable promoter. The expression construct further comprises a transcription initiation site, a transcription termination site, and, within the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcript expressed by the construct preferably comprises translation beginning with a start and stop codon (e.g., UAA, UGA, or UAG) appropriately positioned at the end of the mRNA to be translated, with UAA and UAG being preferred for expression in mammalian or eukaryotic cells.

[0079] The expression vector preferably includes at least one selectable marker, although this is optional. Such markers include, for example, methotrexate (MTX), dihydrofolate reductase (DHFR, U.S. Pat. Nos. 4,399,216, 4,634,665, 4,656,134, 4,956,288, 5,149,636, 5,179,017, ampicillin, neomycin (G418), mycophenolic acid or glutamine synthetase for eukaryotic cell culture. synthetase, GS, U.S. Patent Nos. 5,122,464, 5,770,359, 5,827,739) resistance genes, and tetracycline or ampicillin resistance genes for culture in E. coli and other bacteria or prokaryotes (the above patents are incorporated herein by reference in their entireties). Appropriate culture media and conditions for the above host cells are known in the art. Suitable vectors will be readily apparent to those of skill in the art. Introduction of the vector construct into the host cell can be accomplished by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid mediated transfection, electroporation, transduction, infection or other known methods. Such methods are described in the art, such as Sambrook, supra, Chapters 1-4 and 16-18, and Ausubel, supra, Chapters 1, 9, 13, 15, 16.

[0080] At least one antibody used in the method of the invention may be expressed in modified form, such as a fusion protein, and may contain not only secretion signals, but also additional heterologous functional regions. For example, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the antibody to improve stability and persistence in the host cell during purification or during subsequent processing and storage. Peptide moieties may also be added to the antibody of the invention to facilitate purification. Such regions may be removed prior to final preparation of the antibody or at least one fragment thereof. Such methods are described in many standard laboratory manuals, such as Sambrook, supra, Chapters 17.29-17.42 and 18.1-18.74, Ausubel, supra, Chapters 16, 17 and 18.

[0081] Those skilled in the art are familiar with the numerous expression systems available for the expression of nucleic acids encoding proteins used in the methods of the present invention. Alternatively, the nucleic acids can be expressed in host cells by switching on (by engineering) in the host cells containing endogenous DNA encoding the antibody. Such methods are well known in the art, as described in U.S. Patent Nos. 5,580,734, 5,641,670, 5,733,746, and 5,733,761, which are incorporated herein by reference in their entirety.

[0082] An example of a cell culture useful for the production of an antibody, specified portion or variant thereof is a mammalian cell. Mammalian cell lines are often in the form of a monolayer of cells, although suspensions or bioreactors of mammalian cells can also be used. A number of suitable host cell lines capable of expressing intact glycosylated proteins have been developed in the art, including COS-1 (e.g., ATCC CRL 1650), COS-7 (e.g., ATCC CRL-1651), HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e.g., ATCC CRL1610) and BSC-1 (e.g., ATCC CRL-26) cell lines, Cos-7 cells, CHO cells, hep G2 cells, P3X63Ag8.653, SP2 / 0-Ag14, 293 cells, HeLa cells, and the like, which are readily available, for example, from the American Type Culture Collection, Manassas, Va. (www.atcc.org). Preferred host cells include cells derived from lymphatic system, such as myeloma and lymphoma cells. Particularly preferred host cells are P3X63Ag8.653 cells (ATCC Accession No. CRL-1580) and SP2 / 0-Ag14 cells (ATCC Accession No. CRL-1851). In a particularly preferred embodiment, the recombinant cell is P3X63Ab8.653 or SP2 / 0-Ag14 cell.

[0083] Expression vectors for these cells can include one or more of the following expression control sequences, such as, but not limited to, an origin of replication, a promoter (e.g., the late or early SV40 promoter, a CMV promoter (U.S. Pat. Nos. 5,168,062, 5,385,839), an HSV tk promoter, a pgk (phosphoglycerate kinase) promoter, an EF-1α promoter (U.S. Pat. No. 5,266,491), at least one human immunoglobulin promoter, an enhancer, and / or processing information sites such as ribosome binding sites, RNA splice sites, polyadenylation sites (e.g., the SV40 large T Ag polyaddition site), and transcription termination sequences. See, e.g., Ausubel et al., supra; Sambrook et al., supra. Other cells useful for producing the nucleic acids or proteins of the invention are known and / or available, for example, from the American Type Culture Collection's catalog of cell lines and hybridomas (www.atcc.org) or other known or commercial sources.

[0084] When eukaryotic host cells are used, typically polyadenylation or transcription termination sequences are incorporated into the vector. An example of a termination sequence is the polyadenylation sequence from the bovine growth hormone gene. Sequences for accurate splicing of the transcript can also be included. An example of a splicing sequence is the VP1 intron from SV40 (Sprague, et al., J. Virol. 45:773-781 (1983)). In addition, gene sequences for controlling replication in the host cell can be incorporated into the vector, as is known in the art.

[0085] Antibody purification Anti-IL-23 antibodies can be recovered and purified from recombinant cell cultures by well-known methods, including, but not limited to, protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography. High performance liquid chromatography ("high performance liquid chromatography, HPLC") can also be used for purification. See, e.g., Colligan, Current Protocols in Immunology or Current Protocols in Protein Science, John Wiley & Sons, NY, NY (1997-2001), e.g., chapters 1, 4, 6, 8, 9, 10, each of which is incorporated herein by reference in its entirety.

[0086] Antibodies for use in the methods of the invention include naturally purified products, products of chemical synthetic procedures, and products produced by recombinant methods from eukaryotic hosts, including, for example, yeast, higher plants, insect, and mammalian cells. Depending on the host utilized in the recombinant production procedure, the antibodies may be glycosylated or non-glycosylated, but are preferably glycosylated. Such methods are described in many standard laboratory manuals, such as Sambrook, supra, Sections 17.37-17.42; Ausubel, supra, Chapters 10, 12, 13, 16, 18, and 20; Colligan, Protein Science, supra, Chapters 12-14, all of which are incorporated herein by reference in their entirety.

[0087] Anti-IL-23 antibody The anti-IL-23 antibodies of the present invention comprise at least a portion of an immunoglobulin molecule, such as, but not limited to, at least one ligand binding portion (LBP), such as, but not limited to, a heavy or light chain complementarity determining region (CDR) or a ligand binding portion thereof, a heavy or light chain variable region, a framework region (e.g., FR1, FR2, FR3, FR4, or a fragment thereof, and optionally, including at least one substitution, insertion, or deletion), a heavy or light chain constant region (e.g., at least one CDR or CDR fragment thereof), a heavy or light chain variable region, ... H 1, Hinge 1, Hinge 2, Hinge 3, Hinge 4, C H 2 or C H 3, or a fragment thereof, and optionally including at least one substitution, insertion, or deletion), or any portion thereof. The antibody may include or be derived from any mammal, such as, but not limited to, human, mouse, rabbit, rat, rodent, primate, or any combination thereof.

[0088] The isolated antibodies used in the methods of the invention include the amino acid sequences of the antibodies disclosed herein, encoded by any suitable polynucleotide, or any isolated or prepared antibody. Preferably, the human antibodies or antigen-binding fragments bind to human IL-23, thereby partially or substantially neutralizing at least one biological activity of the protein. Antibodies, or specified portions or variants thereof, that partially or preferably substantially neutralize at least one biological activity of at least one IL-23 protein or fragment, can bind to the protein or fragment, thereby inhibiting an activity mediated through binding of IL-23 to the IL-23 receptor, or through other IL-23-dependent or mediated mechanisms. As used herein, the term "neutralizing antibody" refers to an antibody that can inhibit IL-23-dependent activity by about 20-120%, preferably at least about 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100% or more, depending on the assay. The ability of an anti-IL-23 antibody to inhibit IL-23-dependent activity is preferably assessed by at least one suitable IL-23 protein or receptor assay described herein and / or known in the art. Human antibodies may be of any class (IgG, IgA, IgM, IgE, IgD, etc.) or isotype and may contain kappa or lambda light chains. In one embodiment, the human antibody comprises an IgG heavy chain or defined fragment, e.g., at least one of the following isotypes: IgG1, IgG2, IgG3, or IgG4 (e.g., γ1, γ2, γ3, γ4). Antibodies of this type can be prepared by utilizing transgenic mice or other non-human transgenic mammals that contain at least one human light chain (e.g., IgG, IgA, and IgM) transgene as described herein and / or known in the art. In another embodiment, the anti-IL-23 human antibody comprises an IgG1 heavy chain and an IgG1 light chain.

[0089] The antibody binds to at least one defined epitope specific for at least one IL-23 protein, subunit, fragment, portion, or any combination thereof, which can include at least one antibody binding region that comprises at least a portion of the protein, preferably consisting of at least one extracellular, soluble, hydrophilic, exoportion, or cytoplasmic portion of the protein.

[0090] Generally, a human antibody or antigen-binding fragment comprises an antigen-binding region that comprises at least one human complementarity determining region (CDR1, CDR2, and CDR3) or variant of at least one heavy chain variable region, and at least one human complementarity determining region (CDR1, CDR2, and CDR3) or variant of at least one light chain variable region. The CDR sequences can be derived from human germline sequences or can closely match germline sequences. For example, CDRs derived from synthetic libraries derived from original non-human CDRs can be used. These CDRs can be formed by incorporation of conservative substitutions from original non-human sequences. In another specific embodiment, the antibody or antigen-binding portion or variant can have an antigen-binding region that comprises at least a portion of at least one light chain CDR (i.e., CDR1, CDR2, and / or CDR3) with the corresponding CDR1, 2, and / or 3 amino acid sequences.

[0091] Such antibodies can be prepared by preparing and expressing a nucleic acid molecule (i.e., one or more) encoding the antibody using conventional techniques involving recombinant DNA technology, or by chemically linking together the various portions of the antibody (e.g., CDRs, framework) using conventional techniques, or by using any other suitable methods.

[0092] An anti-IL-23 specific antibody can comprise at least one of a heavy or light chain variable region having a defined amino acid sequence. For example, in a preferred embodiment, an anti-IL-23 antibody comprises at least one of a heavy chain variable region, optionally having the amino acid sequence of SEQ ID NO: 7, and / or at least one of a light chain variable region, optionally having the amino acid sequence of SEQ ID NO: 8. Antibodies that bind to human IL-23 and comprise defined heavy or light chain variable regions can be prepared using suitable methods, such as those employing phage display (Katsube, Y., et al., Int J Mol. Med, 1(5):863-868 (1998)) or transgenic animals, as known in the art and / or described herein. For example, a transgenic mouse comprising a functionally rearranged human immunoglobulin heavy chain transgene and a transgene comprising DNA from a human immunoglobulin light chain locus capable of undergoing functional rearrangement can be immunized with human IL-23 or a fragment thereof to induce the production of antibodies. If desired, antibody-producing cells can be isolated and hybridomas or other immortalized antibody-producing cells can be prepared as described herein and / or known in the art. Alternatively, antibodies, specified portions or variants can be expressed using the encoding nucleic acid, or a portion thereof, in a suitable host cell.

[0093] The present invention also relates to antibodies, antigen-binding fragments, immunoglobulin chains and CDRs that comprise amino acids in sequences that are substantially the same as the amino acid sequences described herein. Preferably, such antibodies or antigen-binding fragments and antibodies that comprise such chains or CDRs have high affinity (e.g., K D About 10 -9M or less). Amino acid sequences that are substantially the same as the sequences described herein include sequences containing conservative amino acid substitutions as well as amino acid deletions and / or insertions. A conservative amino acid substitution refers to the replacement of a first amino acid with a second amino acid that has chemical and / or physical properties (e.g., charge, structure, polarity, hydrophobicity / hydrophilicity) similar to those of the first amino acid. Conservative substitutions include, but are not limited to, replacing one amino acid with another amino acid within the following groups: lysine (K), arginine (R), and histidine (H); aspartate (D) and glutamate (E); asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), K, R, H, D, and E; alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), tryptophan (W), methionine (M), cysteine ​​(C), and glycine (G); F, W, and Y; C, S, and T.

[0094] Amino acid code The amino acids that make up the anti-IL-23 antibodies of the present invention are often abbreviated. Amino acid designations can be provided by designating the amino acid by its one-letter code, its three-letter code, name, or three nucleotide codon(s), and are well understood in the art (see Alberts, B., et al., Molecular Biology of The Cell, Third Ed., Garland Publishing, Inc., New York, 1994).

[0095] [Table 1]

[0096] The anti-IL-23 antibodies used in the methods of the invention may contain one or more amino acid substitutions, deletions, or additions, either by natural mutation or by human manipulation, as specified herein.

[0097] The number of amino acid substitutions that one of skill in the art may make will depend on many factors, including those described above. Generally speaking, the number of amino acid substitutions, insertions or deletions in a given anti-IL-23 antibody, fragment or variant will not exceed 40, 30, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, e.g., 1-30, or any range or value therein, as specified herein.

[0098] Amino acids within an anti-IL-23 specific antibody that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (e.g., Ausubel, supra, Chapters 8, 15; Cunningham and Wells, Science 244:1081-1085 (1989)). The latter procedure introduces single alanine mutations at every residue in the molecule. The resulting substitution mutant molecules are then tested for biological activity, such as, but not limited to, at least one IL-23 neutralizing activity. Sites critical for antibody binding can also be identified by structural analysis, such as crystallization, nuclear magnetic resonance, or photoaffinity labeling (Smith, et al., J. Mol. Biol. 224:899-904 (1992) and de Vos, et al., Science 255:306-312 (1992)).

[0099] The anti-IL-23 antibody can include, but is not limited to, at least one portion, sequence, or combination selected from from five to all of at least one of the contiguous amino acids of SEQ ID NOs: 1, 2, 3, 4, 5, and 6.

[0100] The IL-23 antibody or specific portion or variant may include, but is not limited to, at least one portion, sequence, or combination selected from at least 3 to 5 contiguous amino acids of the above SEQ ID NO:, 5 to 17 contiguous amino acids of the above SEQ ID NO:, 5 to 10 contiguous amino acids of the above SEQ ID NO:, 5 to 11 contiguous amino acids of the above SEQ ID NO:, 5 to 7 contiguous amino acids of the above SEQ ID NO:, or 5 to 9 contiguous amino acids of the above SEQ ID NO:.

[0101] The anti-IL-23 antibody can further optionally comprise at least one polypeptide of 70-100% of the contiguous amino acids of SEQ ID NO: 5, 17, 10, 11, 7, 9, 119, or 108 of the above ..., 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, 9, 119, or 10, 11, 7, Preferably, 70-100% amino acid identity (i.e., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or any range or value therein) is determined using a suitable computer algorithm, as known in the art.

[0102] As known in the art, "identity" is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as determined by the match between such linear sequences. "Identity" and "similarity" can be readily calculated by known methods, including, but not limited to, those described in Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., Siam J. Applied Math., 48:1073 (1988). In addition, percent identity values ​​can be obtained from amino acid and nucleotide sequence alignments generated using default settings in AlignX, a component of Vector NTI Suite 8.0 (Informax, Frederick, MD).

[0103] Preferred methods for determining identity are designed to obtain the highest degree of correspondence between the sequences tested. Methods for determining identity and similarity are codified in publicly available computer programs. Preferred computer program methods for determining identity and similarity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387(1984)), BLASTP, BLASTN, and FASTA (Atschul, SF et al., J.Molec.Biol.215:403-410(1990)). BLAST X programs are publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBINLM NIH Bethesda, Md. 20894: Altschul, S., et al., J. Mol. Biol. 215:403-410 (1990). The well-known Smith Waterman algorithm may also be used to determine identity.

[0104] Preferred parameters for polypeptide sequence comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison matrix: BLOSSUM62 from Hentikoff and Hentikoff, Proc. Natl. Acad. Sci, USA. 89: 10915-10919 (1992) Gap penalty: 12 Gap length penalty: 4 A program useful with these parameters is publicly available as the "Gap" program from the Genetics Computer Group, Madison Wis. The aforementioned parameters are the default parameters for peptide sequence comparisons (as well as no penalty for end gaps).

[0105] Preferred parameters for polynucleotide comparisons include the following: (1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison matrix: match = +10, mismatch = 0 Gap penalty: 50 Gap length penalty: 3 Available as the "Gap" program from the Genetics Computer Group, Madison Wis. These are the default parameters for nucleic acid sequence comparisons.

[0106] By way of example, a polynucleotide sequence may be identical to another sequence, i.e., 100% identical, or may contain up to a certain integer number of nucleotide changes compared to a reference sequence. Such changes may be selected from the group consisting of at least one nucleotide deletion, substitution (including transitions and transversions), or insertion, and the changes may occur at the 5' or 3' terminal position of the reference nucleotide sequence, or anywhere between these terminal positions, and may be interspersed between nucleotides of the reference sequence, either individually, or in one or more adjacent groups within the reference sequence. The number of nucleotide changes may be determined by multiplying the total number of nucleotides in the sequence by the numerical percentage of the corresponding percent identity (divided by 100) and subtracting the product from the total number of nucleotides in the sequence, or is determined by n.sub.n.ltorsim.x.sub.n-(x.sub.ny), where n.sub.n is the number of nucleotide changes, x.sub.n is the total number of nucleotides in the sequence, and y is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95%, etc., and any non-integer product of x.sub.n and y is rounded down to the nearest integer before subtraction from x.sub.n.

[0107] The modification of the polynucleotide sequence encoding the SEQ ID NO can create nonsense, missense, or frameshift mutations in the coding sequence, thereby modifying the polypeptide encoded by the polynucleotide after such modification. Similarly, the polypeptide sequence can be identical to the reference sequence of the SEQ ID NO, i.e., 100% identical, or can contain up to a certain integer number of amino acid modifications compared to the reference sequence such that the percent identity is less than 100%. Such modifications are selected from the group consisting of deletion, substitution (including conservative and non-conservative substitution), or insertion of at least one amino acid, and the modifications can occur at the amino or carboxy terminal position of the reference polypeptide sequence or anywhere between these terminal positions, and can be interspersed between the amino acids of the reference sequence either individually or in one or more adjacent groups within the reference sequence. The number of amino acid modifications for a given identity percentage can be calculated by multiplying the total number of amino acids in the SEQ ID NO by the numerical percent of the corresponding identity percentage (divided by 100) and subtracting the product from the total number of amino acids in the SEQ ID NO, or is determined by n.sub.a.ltorsim.x.sub.a-(x.sub.ay), where n.sub.a is the number of amino acid changes, x.sub.a is the total number of amino acids in the sequence ID above, and y is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, etc., and any non-integer product of x.sub.a and y is rounded down to the nearest integer before subtraction from x.sub.a.

[0108] Exemplary heavy and light chain variable region sequences, and portions thereof, are set forth in the SEQ ID NOs. An antibody of the invention, or a specified variant thereof, may comprise any number of contiguous amino acid residues from an antibody of the invention, the number being selected from the group of integers consisting of 10-100% of the number of contiguous residues in an anti-IL-23 antibody. Optionally, this subsequence of contiguous amino acids is at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, or more amino acids in length, or any range or value therein. Additionally, the number of such subsequences may be any integer selected from the group consisting of 1-20, such as at least 2, 3, 4, or 5.

[0109] As will be apparent to one of skill in the art, the present invention includes at least one biologically active antibody of the present invention. A biologically active antibody has a specific activity that is at least 20%, 30%, or 40%, and preferably at least 50%, 60%, or 70%, and most preferably at least 80%, 90%, or 95% to 100% or more (including but not limited to up to 10-fold the specific activity) of that of a natural (non-synthetic), endogenous, or related and known antibody. Methods for assaying and quantitatively measuring enzymatic activity and substrate specificity are well known to those of skill in the art.

[0110] In another aspect, the invention relates to human antibodies and antigen-binding fragments described herein that are modified by the covalent attachment of an organic moiety. Such modifications can generate antibodies or antigen-binding fragments with improved pharmacokinetic properties (e.g., increased serum half-life in vivo). The organic moiety can be a linear or branched hydrophilic polymer group, a fatty acid group, or a fatty acid ester group. In certain embodiments, the hydrophilic polymer group can be a polyalkane glycol (e.g., polyethylene glycol (PEG), polypropylene glycol (PPG)), a carbohydrate polymer, an amino acid polymer, or polyvinylpyrrolidone, with a molecular weight of about 800 to about 120,000 daltons, and the fatty acid group or fatty acid ester group can contain about 8 to about 40 carbon atoms.

[0111] The modified antibodies and antigen-binding fragments may contain one or more organic moieties that are directly or indirectly covalently attached to the antibody. Each organic moiety attached to the antibody or antigen-binding fragment of the present invention may be independently a hydrophilic polymer group, a fatty acid group, or a fatty acid ester group. As used herein, the term "fatty acid" includes monocarboxylic and dicarboxylic acids. As used herein, the term "hydrophilic polymer group" refers to an organic polymer that is more soluble in water than octane. For example, polylysine is more soluble in water than octane. Thus, antibodies modified by the covalent attachment of polylysine are encompassed by the present invention. Suitable hydrophilic polymers for modifying the antibodies of the present invention may be linear or branched, and include, for example, polyalkane glycols (e.g., PEG, monomethoxy-polyethylene glycol (mPEG), PPG, etc.), carbohydrates (e.g., dextran, cellulose, oligosaccharides, polysaccharides, etc.), polymers of hydrophilic amino acids (e.g., polylysine, polyarginine, polyaspartic acid, etc.), polyalkane oxides (e.g., polyethylene oxide, polypropylene oxide, etc.), and polyvinylpyrrolidone. Preferably, the hydrophilic polymers for modifying the antibodies of the present invention have a molecular weight of about 800 to about 150,000 daltons as individual molecular entities. For example, PEG 5000 and PEG 20,000 can be used, where the subscript is the average molecular weight of the polymer in Daltons. The hydrophilic polymer group can be substituted with 1 to about 6 alkyl groups, fatty acid groups, or fatty acid ester groups. Hydrophilic polymers substituted with fatty acid or fatty acid ester groups can be prepared by utilizing a suitable method. For example, a polymer containing an amine group can be linked to a carboxylate of a fatty acid or fatty acid ester, and an activated carboxylate on the fatty acid or fatty acid ester (e.g., activated with N,N-carbonyldiimidazole) can be linked to a hydroxyl group on the polymer.

[0112] Fatty acids and fatty acid esters suitable for modifying antibodies of the invention may be saturated or may contain one or more units of unsaturation. Fatty acids suitable for modifying antibodies of the invention include, for example, n-dodecanoate (C 12 , laurate), n-tetradecanoate (C 14 , myristate), n-octadecanoate (C 18 , stearate), n-eicosanoate (C 20 , arachidate), n-docosanoate (C 22 , behenate), n-triacontanoate (C 30 ), n-tetracontanoate (C 40 ), cis-Δ9-octadecanoate (C 18 , oleate), all cis-Δ5,8,11,14-eicosatetraenoate (C 20 , arachidonate), octanedioic acid, tetradecanedioic acid, octadecanedioic acid, docosanedioic acid, and the like. Suitable fatty acid esters include monoesters of dicarboxylic acids containing a straight or branched chain lower alkyl group. The lower alkyl group may contain from 1 to about 12, preferably from 1 to about 6, carbon atoms.

[0113] Modified human antibodies and antigen-binding fragments can be prepared using suitable methods, such as by reaction with one or more modifying agents. As used herein, the term "modifying agent" refers to a suitable organic group (e.g., hydrophilic polymer, fatty acid, fatty acid ester) that includes an activating group. An "activating group" is a chemical moiety or functional group that can react with a second chemical group under appropriate conditions, thereby forming a covalent bond between the modifying agent and the second chemical group. For example, amine-reactive activating groups include electrophilic groups such as tosylate, mesylate, halo (chloro, bromo, fluoro, iodo), N-hydroxysuccinimidyl esters (NHS), and the like. Activating groups that can react with thiols include, for example, maleimide, iodoacetyl, acrylolyl, pyridyl disulfide, 5-thiol-2-nitrobenzoic acid thiol (TNB-thiol), and the like. Aldehyde functional groups can be linked to amine- or hydrazide-containing molecules, and azide groups can react with trivalent phosphorus groups to form phosphoramidate or phosphorimide bonds. Suitable methods for introducing activating groups into molecules are known in the art (see, for example, Hermanson, GT, Bioconjugate Techniques, Academic Press: San Diego, CA (1996)). Activating groups can be attached directly to organic groups (e.g., hydrophilic polymers, fatty acids, fatty acid esters) or to linker moieties, such as divalent C1-C 12The linker moiety may be linked via a group (wherein one or more carbon atoms may be replaced with a heteroatom such as oxygen, nitrogen, or sulfur). Suitable linker moieties include, for example, tetraethylene glycol, -(CH2)3-, -NH-(CH2)6-NH-, -(CH2)2-NH-, and -CH2-O-CH2-CH2-O-CH2-CH2-O-CH-NH-. Modifiers containing a linker moiety can be generated by forming an amide bond between a free amine and a fatty acid carboxylate, for example, by reacting a mono-Boc-alkyldiamine (e.g., mono-Boc-ethylenediamine, mono-Boc-diaminohexane) with a fatty acid in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). The Boc protecting group can be removed from the product by treatment with trifluoroacetic acid (TFA) to expose a primary amine that can be coupled to another carboxylate as described, or it can be reacted with maleic anhydride and the resulting product cyclized to generate an activated maleimide derivative of a fatty acid (see, for example, Thompson et al., International Publication No. WO 92 / 16221, the teachings of which are incorporated herein by reference in their entirety).

[0114] Modified antibodies can be produced by reacting a human antibody or antigen-binding fragment with a modifying agent. For example, an organic moiety can be attached to the antibody in a non-site-specific manner using an amine-reactive modifying agent, e.g., an NHS ester of PEG. Modified human antibodies or antigen-binding fragments can also be prepared by reducing disulfide bonds (e.g., intrachain disulfide bonds) of an antibody or antigen-binding fragment. The reduced antibody or antigen-binding fragment can then be reacted with a thiol-reactive modifying agent to produce the modified antibody of the invention. Modified human antibodies and antigen-binding fragments containing organic moieties attached to specific sites of the antibodies of the invention can be prepared using suitable methods, such as reverse proteolysis (Fisch et al., Bioconjugate Chem., 3:147-153 (1992); Werlen et al., Bioconjugate Chem., 5:411-417 (1994); Kumaran et al., Protein Sci. 6(10):2233-2241 (1997); Itoh et al., Bioorg. Chem., 24(1):59-68 (1996); Capellas et al., Biotechnol. Bioeng., 56(4):456-463 (1997)) and those described in Hermanson, GT, Bioconjugate Techniques, Academic Press: San Diego, CA (1996).

[0115] The methods of the invention also employ anti-IL-23 antibody compositions comprising at least one, at least two, at least three, at least four, at least five, at least six or more anti-IL-23 antibodies provided in a non-naturally occurring composition, mixture or form as described herein and / or known in the art. Such compositions include non-naturally occurring compositions comprising at least one or two full-length, C- and / or N-terminal deletion variants, domains, fragments or specified variants of anti-IL-23 antibody amino acid sequences selected from the group consisting of 70-100% of the contiguous amino acids of said sequences, or specified fragments, domains or variants thereof. A preferred anti-IL23 antibody composition comprises at least one or two full-length, fragments, domains or variants as at least one CDR or LBP-containing portion of an anti-IL23 antibody sequence described herein, or specified fragments, domains or variants thereof, for example, 70-100% of the above SEQ ID NOs. More preferred compositions comprise, for example, 70-100% of the above SEQ ID NOs, or 40-99% of at least one of the specified fragments, domains, or variants thereof. Such composition percentages may be by weight, volume, concentration, molarity, or molar concentrations as liquid or dry solutions, mixtures, suspensions, emulsions, particles, powders, or colloids, as known in the art or as described herein.

[0116] Antibody compositions containing further therapeutically active ingredients The antibody compositions used in the methods of the present invention can optionally further comprise an effective amount of at least one compound or protein selected from at least one of the following: anti-infective agents, cardiovascular (CV) agents, central nervous system (CNS) agents, autonomic nervous system (ANS) agents, respiratory agents, gastrointestinal (GI) tract agents, hormonal agents, fluid or electrolyte balancing agents, hemodynamic agents, antineoplastic agents, immunomodulatory agents, ophthalmic, otic or nasal agents, topical agents, nutritional agents, etc. Such agents are well known in the art (see, e.g., Nursing 2001 Handbook of Drugs, 21), including their respective formulations, indications, dosages, and administration as set forth herein. st edition, Springhouse Corp., Springhouse, PA, 2001; Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ; Pharmacology Handbook, Wells et al., Appleton & Lange, Stamford, CT, each of which is incorporated herein by reference).

[0117] As examples of drugs that can be combined with the antibodies of the method of the present invention, the anti-infective drug can be at least one selected from amebicide or at least one antiprotozoal drug, anthelmintic drug, antifungal drug, antimalarial drug, antituberculous drug or at least one antilepromycotic drug, aminoglycoside, penicillin, cephalosporin, tetracycline, sulfonamide, fluoroquinolone, antiviral drug, macrolide anti-infective drug, and miscellaneous anti-infective drugs. The hormonal drug can be at least one selected from corticosteroid, androgen, or at least one anabolic steroid, estrogen, or at least one progestin, gonadotropin, antidiabetic drug, or at least one glucagon, thyroid hormone, thyroid hormone antagonist, pituitary hormone, and parathyroid mimetic drug. The at least one cephalosporin can be at least one selected from cefaclor, cefadroxil, cefazolin sodium, cefdinir, cefepime hydrochloride, cefixime, cefmetazole sodium, cefonicid sodium, cefoperazone sodium, cefotaxime sodium, cefotetan disodium, cefoxitin sodium, cefpodoxime proxetil, cefprozil, ceftazidime, ceftibuten, ceftizoxime sodium, ceftriaxone sodium, cefuroxime axetil, cefuroxime sodium, cephalexin hydrochloride, cephalexin monohydrate, cephradine, and loracarbef.

[0118] The at least one corticosteroid can be at least one selected from betamethasone, betamethasone acetate or betamethasone sodium phosphate, betamethasone sodium phosphate, cortisone acetate, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, fludrocortisone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone cypionate, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, methylprednisolone, methylprednisolone acetate, methylprednisolone sodium succinate, prednisolone, prednisolone acetate, prednisolone sodium phosphate, prednisolone tebutate, prednisone, triamcinolone, triamcinolone acetonide, and triamcinolone diacetate. The at least one androgenic or anabolic steroid can be at least one selected from danazol, fluoxymesterone, methyltestosterone, nandrolone decanoate, nandrolone phenpropionate, testosterone, testosterone cypionate, testosterone enanthate, testosterone propionate, and testosterone transdermals.

[0119] The at least one immunosuppressant can be at least one selected from azathioprine, basiliximab, cyclosporine, daclizumab, lymphocyte immunoglobulin, muromonab-CD3, mycophenolate mofetil, mycophenolate mofetil hydrochloride, sirolimus, and tacrolimus.

[0120] The at least one topical anti-infective agent can be at least one selected from acyclovir, amphotericin B, azelaic acid cream, bacitracin, butoconazole nitrate, clindamycin phosphate, clotrimazole, econazole nitrate, erythromycin, gentamicin sulfate, ketoconazole, mafenide acetate, metronidazole (topical), miconazole nitrate, mupirocin, naftifine hydrochloride, neomycin sulfate, nitrofurazone, nystatin, silver sulfadiazine, terbinafine hydrochloride, terconazole, tetracycline hydrochloride, tioconazole, and tolnaftate. The at least one scabicide or pediculicide can be at least one selected from crotamiton, lindane, permethrin, and pyrethrins. The at least one topical corticosteroid can be at least one selected from betamethasone dipropionate, betamethasone valerate, clobetasol propionate, desonide, desoximetasone, dexamethasone, dexamethasone sodium phosphate, diflorasone diacetate, fluocinolone acetonide, fluocinonide, flurandrenolide, fluticasone propionate, halcionide, hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone valerate, mometasone furoate, and triamcinolone acetonide. (See, e.g., pages 1098-1136 of Nursing 2001 Drug Handbook.)

[0121] Anti-IL-23 antibody compositions comprise at least one anti-IL-23 antibody that is contacted or administered to a cell, tissue, organ, animal, or patient in need of such modulation, treatment, or therapy, and optionally further comprise at least one TNF antagonist (e.g., but not limited to, a TNF chemical or protein antagonist, a TNF monoclonal or polyclonal antibody or fragment, a soluble TNF receptor (e.g., p55, p70, or p85) or fragment, a fusion polypeptide thereof, or a small molecule TNF antagonist, such as TNF binding protein I or II (TBP-1 or TBP-II), nerelimonomab, infliximab, or erythropoietin (ER) (TNF-α, T ... The composition may further include at least one of any suitable and effective amount of compositions or pharmaceutical compositions including at least one selected from the group consisting of antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, etanercept, aurothiomalate, hydroxychloroquine sulfate, leflunomide, sulfasalazine), immunizing agents, immunoglobulins, immunosuppressants (e.g., basiliximab, cyclosporine, daclizumab), cytokines, or cytokine antagonists. Non-limiting examples of such cytokines include, but are not limited to, any of IL-1 to IL-23 (e.g., IL-1, IL-2, etc.). Suitable dosages are well known in the art. For example, see Wells et al., eds., Pharmacotherapy Handbook, 2 nd Edition, Appleton and Lange, Stamford, CT (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, CA (2000), each of which is incorporated herein by reference in its entirety.

[0122] The anti-IL-23 antibody compound, composition, or mixture used in the method of the present invention may further comprise at least one of any suitable auxiliary agent, such as, but not limited to, a diluent, a binder, a stabilizer, a buffer, a salt, a lipophilic solvent, a preservative, an adjuvant, etc. Pharmaceutically acceptable auxiliary agents are preferred. Non-limiting examples of and methods for preparing such sterile solutions are well known in the art and are described, for example, in Gennaro, Ed., Remington's Pharmaceutical Sciences, 1896, 1999, 1998, 19 ...0, 1990, 1990, 1990, 1991, 1992, 1992, 1993, 1994, 1995, 1996, 1997, 1999, 1990, 1991, 1992, 1993, 1994, 1995, 1996, 1997, 1999, 1999, 19 th Edition, Mack Publishing Co. (Easton, PA), 1990. As is well known in the art or described herein, pharma- ceutically acceptable carriers suitable for the mode of administration, solubility, and / or stability of the anti-IL-23 antibody, fragment, or variant composition can be routinely selected.

[0123] Pharmaceutical excipients and additives useful in the present compositions include, but are not limited to, proteins, peptides, amino acids, lipids, and carbohydrates (e.g., saccharides including monosaccharides, disaccharides, trisaccharides, tetrasaccharides, and oligosaccharides, derivatized sugars such as alditols, aldonic acids, esterified sugars, and polysaccharides or sugar polymers), which may be present alone or in combination and comprise 1-99.99% by weight or volume, alone or in combination. Exemplary protein excipients include serum albumins such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acids / antibody components that may also function in a buffering capacity include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. One preferred amino acid is glycine.

[0124] Carbohydrate excipients suitable for use in the present invention include, for example, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, etc., disaccharides such as lactose, sucrose, trehalose, cellobiose, etc., polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starches, etc., alditols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), myo-inositol, etc. Preferred carbohydrate excipients for use in the present invention are mannitol, trehalose, and raffinose.

[0125] The anti-IL-23 antibody composition may also include a buffer or pH adjusting agent, typically a salt prepared from an organic acid or base. Representative buffers include organic acid salts such as salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid, Tris, tromethamine hydrochloride, or phosphate buffers. A preferred buffer for use in the composition is an organic acid salt such as citric acid.

[0126] In addition, anti-IL-23 antibody compositions may include polymeric excipients / additives such as polyvinylpyrrolidone, Ficoll (a polymeric sugar), dextrates (e.g., cyclodextrins such as 2-hydroxypropyl-β-cyclodextrin), polyethylene glycol, flavorings, antimicrobials, sweeteners, antioxidants, antistatic agents, surfactants (e.g., polysorbates such as "TWEEN 20" and "TWEEN 80"), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0127] These and additional known pharmaceutical excipients and / or additives suitable for use in the anti-IL-23 antibody, portion or variant compositions according to the invention are known in the art and are described, for example, in "Remington: The Science & Practice of Pharmacy," 1999. thed., Williams&Williams, (1995) and “Physician's Desk Reference”, 52 nd ed., Medical Economics, Montvale, NJ (1998), the disclosures of which are incorporated herein by reference in their entireties. Preferred carrier or excipient materials are carbohydrates (e.g., monosaccharides and alditols) and buffers (e.g., citric acid) or polymeric agents. Exemplary carrier molecules are mucopolysaccharides, hyaluronic acid, which may be useful for intra-articular delivery.

[0128] formulation As mentioned above, the present invention provides stable formulations, preferably saline or phosphate buffer with selected salts, as well as preservative-containing preservative solutions and formulations, and versatile preservative formulations suitable for pharmaceutical or veterinary use, comprising at least one anti-IL23 antibody in a pharma- ceutically acceptable formulation. The preservative formulations contain at least one known preservative, optionally selected from the group consisting of at least one phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate), alkylparaben (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof, in an aqueous diluent. As known in the art, the range of concentrations may be from 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 may be used.Non-limiting examples include no preservatives, 0.1-2% m-cresol (e.g., 0.2, 0.3, 0.4, 0.5, 0.9, 1.0%), 0.1-3% benzyl alcohol (e.g., 0.5, 0.9, 1.1, 1.5, 1.9, 2.0, 2.5%), 0.001-0.5% thimerosal (e.g., 0.005, 0.01), 0.001-2.0% phenol (e.g., 0. 0.05, 0.25, 0.28, 0.5, 0.9, 1.0%), 0.0005-1.0% alkylparaben(s) (e.g., 0.00075, 0.0009, 0.001, 0.002, 0.005, 0.0075, 0.009, 0.01, 0.02, 0.05, 0.075, 0.09, 0.1, 0.2, 0.3, 0.5, 0.75, 0.9, 1.0%), and the like.

[0129] As discussed above, the methods of the invention employ articles of manufacture comprising packaging and at least one vial containing a solution of at least one anti-IL-23 specific antibody, optionally with a buffer and / or preservative formulated in an aqueous diluent, the packaging comprising a label indicating that such solution can be maintained for 1, 2, 3, 4, 5, 6, 9, 12, 18, 20, 24, 30, 36, 40, 48, 54, 60, 66, 72 hours or more. The invention further employs articles of manufacture comprising packaging and a first vial containing a lyophilized anti-IL-23 specific antibody and a second vial containing an aqueous diluent of the formulated buffer or preservative, the packaging comprising a label instructing a patient to reconstitute the anti-IL-23 specific antibody with the aqueous diluent to form a solution that can be maintained for 24 hours or more.

[0130] The anti-IL-23 specific antibodies used in accordance with the present invention may be produced by recombinant means, including production from mammalian cells or transgenic preparations, or may be purified from other biological sources, as described herein or known in the art.

[0131] Ranges of anti-IL-23 specific antibodies include those amounts that will give a concentration of about 1.0 μg / mL to about 1000 mg / mL upon reconstitution for wet / dry systems, although lower and higher concentrations are workable and depend on the intended delivery vehicle, e.g., for solution formulations, as opposed to transdermal patch, pulmonary, transmucosal, or osmotic or micropump methods.

[0132] Preferably, the aqueous diluent optionally further comprises a pharma- ceutically acceptable preservative. Preferred preservatives include those selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or mixtures thereof. The concentration of the preservative used in the formulation is sufficient to produce an antimicrobial effect. Such concentration will vary depending on the preservative selected and can be easily determined by one skilled in the art.

[0133] Other excipients, such as isotonicity agents, buffers, antioxidants, and preservative enhancers, can be optionally and preferably added to the diluent. An isotonicity agent, such as glycerin, is commonly used at a known concentration. Preferably, a physiologically tolerable buffer is added to provide improved pH control. The formulation can cover a wide range of pH, such as from about pH 4 to about pH 10, and preferably from about pH 5 to about pH 9, and most preferably from about 6.0 to about 8.0. Preferably, the 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 phosphate buffered saline (PBS).

[0134] Other additives, such as pharma- ceutically acceptable solubilizers, such as Tween 20 (polyoxyethylene (20) sorbitan monolaurate), Tween 40 (polyoxyethylene (20) sorbitan monopalmitate), Tween 80 (polyoxyethylene (20) sorbitan monooleate), Pluronic F68 (polyoxyethylene polyoxypropylene block copolymer), and PEG (polyethylene glycol), or non-ionic surfactants, such as polysorbate 20 or 80, or poloxamer 184 or 188, Pluronic® polyls, other block copolymers, and chelating agents, such as EDTA and EGTA, can be optionally added to the formulation or composition to reduce aggregation. These additives are particularly useful when pumps or plastic containers are used to administer the formulation. The presence of pharma-ceutically acceptable surfactants reduces the tendency of proteins to aggregate.

[0135] The formulations can be prepared by a process that includes mixing at least one anti-IL-23 specific antibody with a preservative selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, alkyl parabens (methyl, ethyl, propyl, butyl, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal or mixtures thereof in an aqueous diluent. Mixing at least one anti-IL-23 specific antibody with a preservative in an aqueous diluent is performed using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a quantity of at least one anti-IL-23 specific antibody in a buffer solution is combined with a desired preservative in a sufficient amount of buffer solution to provide the desired concentration of protein and preservative. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of the components, the use or non-use of additional additives, the temperature and pH during preparation of the formulation are all factors that can be optimized for the administration concentration and administration means used.

[0136] The formulations can be provided to patients as clear solutions or as dual vials containing a vial of lyophilized anti-IL-23 specific antibody reconstituted with a second vial containing water, preservatives and / or excipients, preferably phosphate buffer and / or saline, and the selected salt in an aqueous diluent. Either the single solution vial or the dual vial requiring reconstitution can be reused multiple times to satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0137] The product is useful for administration over a period ranging from immediate to 24 hours or more. Thus, the products claimed by the present invention provide significant benefits to patients. The formulations of the present invention can be safely stored, optionally at temperatures between about 2° C. and about 40° C., and retain the biological activity of the protein for extended periods, thus allowing the packaging label to indicate that the solution can be retained and / or used for periods of 6, 12, 18, 24, 36, 48, 72, or 96 hours or more. In cases where a preserved diluent is used, such labeling can include use up to 1-12 months, half a year, one and a half years, and / or up to two years.

[0138] A solution of an anti-IL-23 specific antibody can be prepared by a process that includes mixing at least one antibody in an aqueous diluent. The mixing is carried out using conventional dissolution and mixing procedures. To prepare a suitable diluent, for example, a quantity of at least one antibody in water or a buffer is combined in an amount sufficient to provide the desired concentration of protein, and optionally a preservative or buffer. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of the components, whether or not additional additives are used, the temperature and pH at which the formulation is prepared are all factors that can be optimized for the administration concentration and administration means used.

[0139] The claimed products can be provided to patients as clear solutions or as combination vials containing a vial of at least one lyophilized anti-IL-23 specific antibody that is reconstituted with a second vial containing an aqueous diluent. Either the single solution vial or the dual vial requiring reconstitution can be reused multiple times to satisfy single or multiple patient treatment cycles, thus providing a more convenient treatment regimen than is currently available.

[0140] The claimed products can be provided indirectly to patients by providing a pharmacy, clinic, or other such institution or facility with a combination vial containing a clear solution or a vial of at least one lyophilized anti-IL-23 specific antibody reconstituted with a second vial containing an aqueous diluent, where the clear solution can be up to a liter or even more in volume, from which smaller amounts of the at least one antibody solution can be removed one or more times from the larger container and transferred to smaller vials and provided to customers and / or patients by the pharmacy or clinic.

[0141] Approved devices that contain single vial systems include BD Pens, BD Autojector®, Humaject®, NovoPen®, BD® Pen, AutoPen®, and pen-type injector devices for solution delivery such as OptiPen®, GenotropinPen®, Genotronorm Pen®, Humatro Pen®, Reco-Pen®, Roferon Pen®, Biojector®, Iject®, J-tip Needle-Free Injector®, Intraject®, Medi-Ject®, and Smartject® (e.g., from Becton Dickensen (Franklin Lakes, NJ, www.bectondickenson.com), Disetronic (Burgdorf, Switzerland, www.disetronic.com), Bioject, Portland, Oregon (www.bioject.com), National Suitable devices include those manufactured or developed by Medical Products, Weston Medical (Peterborough, UK, www.weston-medical.com), Medi-Ject Corp (Minneapolis, MN, www.mediject.com), and similar suitable devices. Approved devices that include dual vial systems include pen-type injector systems for reconstituting lyophilized medications in cartridges for delivery of the reconstituted solution, such as HumatroPen®. Examples of other suitable devices include pre-filled syringes, auto-injectors, needleless syringes, and needleless IV infusion sets.

[0142] The article of manufacture may include packaging material. The packaging material provides the conditions under which the product may be used, in addition to any information required by regulatory agencies. The packaging material of the present invention provides instructions to the patient, if applicable, to reconstitute at least one anti-IL-23 antibody with an aqueous diluent to form a solution and use this solution for a period of 2-24 hours or more in a wet / dry two vial product. In the case of a single vial solution product, pre-filled syringe, or auto-injector, the label indicates that such solution may be used for a period of 2-24 hours or more. The article of manufacture is useful for human pharmaceutical product applications.

[0143] The formulations used in the methods of the present invention can be prepared by a process that includes mixing an anti-IL-23 antibody and a selected buffer, preferably saline or a phosphate buffer containing a selected salt. Mixing the anti-IL-23 antibody and the buffer in an aqueous diluent is performed using conventional dissolution and mixing procedures. To prepare a suitable formulation, for example, a quantity of at least one antibody in water or buffer is combined with a desired buffer in a sufficient amount of water to provide the desired concentration of protein and buffer. Variations of this process will be recognized by those skilled in the art. For example, the order of addition of the components, whether or not additional additives are used, the temperature and pH at which the formulation is prepared are all factors that can be optimized for the administration concentration and administration means used.

[0144] The method of the present invention provides pharmaceutical compositions including various formulations useful and acceptable for administration to human or animal patients. Such pharmaceutical compositions are prepared using "standard conditions" water as a diluent and routine methods well known to those skilled in the art. For example, buffer components such as histidine and histidine monohydrochloride hydrate may be provided first, followed by the addition of an appropriate non-final volume of "standard conditions" water diluent, sucrose, and polysorbate 80. The isolated antibody may then be added. Finally, the volume of the pharmaceutical composition is adjusted to the desired final volume under "standard conditions" using water as a diluent. Those skilled in the art will recognize several other methods suitable for the preparation of pharmaceutical compositions.

[0145] The pharmaceutical composition may be an aqueous solution or suspension containing the indicated mass of each component per volume unit of water or having the indicated pH at "standard conditions". As used herein, the term "standard conditions" refers to a temperature of 25°C + / - 2°C and a pressure of 1 atmosphere. The term "standard conditions" is not used in the art to refer to a single art recognized set of temperatures or pressures, but is instead a reference condition that specifies the temperature and pressure used to describe a solution or suspension containing a particular composition under reference "standard conditions" conditions. This is because the volume of a solution is in part a function of temperature and pressure. Those skilled in the art will recognize that pharmaceutical compositions equivalent to those disclosed herein may be produced at other temperatures and pressures. The equivalence of such pharmaceutical compositions to those disclosed herein should be determined under "standard conditions" conditions defined above (e.g., 25°C + / - 2°C and a pressure of 1 atmosphere).

[0146] Importantly, such pharmaceutical compositions may contain a component mass of "about" a certain value (e.g., "about 0.53 mg L-histidine") or have a pH value of about a certain value per unit volume of the pharmaceutical composition. The component mass or pH value present in the pharmaceutical composition is "about" a given numerical value when the isolated antibody is present in the pharmaceutical composition or when the isolated antibody is present in the pharmaceutical composition after it is removed from the pharmaceutical composition (e.g., by dilution) and can bind to a peptide chain. That is, the value, such as the component mass value or pH value, is "about" a given numerical value when the binding activity of the isolated antibody is maintained and detectable after placing the isolated antibody in a pharmaceutical composition.

[0147] Competitive binding analysis is performed to determine whether IL-23 specific mAbs bind similar or different epitopes and / or compete with each other. Abs are individually coated onto ELISA plates. Competing mAbs are added, followed by biotinylated hrIL-23. For a positive control, the same mAb for coating can be used as a competing mAb ("self-competition"). IL-23 binding is detected using streptavidin. These results indicate whether the mAbs recognize similar or partially overlapping epitopes on IL-23.

[0148] One aspect of the methods of the invention involves administering a pharmaceutical composition to a patient.

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

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

[0151] Other formulations or methods for stabilizing anti-IL-23 antibodies may be other than clear solutions of lyophilized powders containing the antibodies. Non-clear solutions include formulations containing particulate suspensions, which are compositions containing anti-IL-23 antibodies in structures of various sizes known variously as microspheres, microparticles, nanoparticles, nanospheres, or liposomes. Such relatively homogeneous essentially spherical particulate formulations containing active agents can be formed by contacting an aqueous phase containing active agents and polymers with a non-aqueous phase, as taught in U.S. Pat. No. 4,589,330, and then evaporating the non-aqueous phase to cause coalescence of particles from the aqueous phase. Porous microparticles can be prepared by using a first phase containing active agents and polymers dispersed in a continuous solvent, and removing the solvent from the suspension by lyophilization or dilution-extraction-precipitation, as taught in U.S. Pat. No. 4,818,542. Preferred polymers for such preparations are gelatin agar, starch, arabinogalactan, albumin, collagen, polyglycolic acid, polylactic acid, glycolide-L(-)lactide, poly(epsilon-caprolactone, poly(epsilon-caprolactone-co-lactic acid), poly(epsilon-caprolactone-co-glycolic acid), poly(β-hydroxybutyric acid), polyethylene oxide, polyethylene, poly(alkyl-2-cyanoacrylate), poly(hydroxyethyl methacrylate), polyamide, poly(amino acids), poly(2-hydroxyethyl DL-aspartamide), poly(ester urea), poly(L-phenylalanine / ethylene glycol), poly( ... The preferred polymers are natural or synthetic copolymers or polymers selected from the group consisting of poly(methyl methacrylate), poly(ethylene glycol / 1,6-diisocyanatohexane), and poly(methyl methacrylate). Particularly preferred polymers are polyesters such as polyglycolic acid, polylactic acid, glycolide-L(-)lactide, poly(epsilon-caprolactone), poly(epsilon-caprolactone-co-lactic acid), and poly(epsilon-caprolactone-co-glycolic acid). Solvents useful for dissolving the polymer and / or active agent include water, hexafluoroisopropanol, methylene chloride, tetrahydrofuran, hexane, benzene, or hexafluoroacetone sesquihydrate.The process of dispersing the active-containing phase into the second phase may include forcing said first phase under pressure through an orifice in a nozzle to affect droplet formation.

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

[0153] Anti-IL-23 antibodies in any of the stable or preserved formulations or solutions described herein can be administered to a patient according to the invention via a variety of delivery methods, as known in the art, such as SC or IM injection, transdermal, transpulmonary, transmucosal, implants, osmotic pumps, cartridges, micropumps, or other means well known in the art and understood by the skilled artisan.

[0154] therapeutic application The present invention also provides methods for modulating or treating psoriasis in a cell, tissue, organ, animal, or patient using at least one IL-23 antibody of the present invention, e.g., by administering to or contacting the cell, tissue, organ, animal, or patient with a therapeutically effective amount of an IL-23 specific antibody, as known in the art or described herein.

[0155] Any of the methods of the invention may comprise administering to a cell, tissue, organ, animal, or patient in need of such modulation, treatment, or therapy an effective amount of a composition or pharmaceutical composition comprising an anti-IL-23 antibody. Such methods may optionally further comprise co-administration or combination therapy for the treatment of such disease or disorder, wherein administering the at least one anti-IL-23 antibody, specified portion, or variant thereof, is in combination with at least one TNF antagonist (such as, but not limited to, a chemical or proteinaceous TNF antagonist, a TNF monoclonal or polyclonal antibody or fragment, a soluble TNF receptor (e.g., p55, p70, or p85) or fragment, a fusion polypeptide thereof, or a small molecule TNF antagonist, such as TNFR2, TNFR3, TNFR4, TNFR5, TNFR6, TNFR7, TNFR8, TNFR9, TNFR10, TNFR11, TNFR12, TNFR13, TNFR14, TNFR15, TNFR16, TNFR17, TNFR18, TNFR19, TNFR20, TNFR21, TNFR22, TNFR23, TNFR24, TNFR25, TNFR26, TNFR27, TNFR28, TNFR29, TNFR30, TNFR31, TNFR32, TNFR33, TNFR41, TNFR42, TNFR53, TNFR54, TNFR55, TNFR55, TNFR55, TNFR61, TNFR72, TNFR83, TNFR94, TNFR105, TNFR116, TNFR126, TNFR14, TNFR155, TNFR156, TNFR165, TNFR176, TNFR187, TNFR195, TNFR196, F-binding protein I or II (TBP-1 or TBP-II), nerelimomab, infliximab, etanercept (Enbrel™), adalimumab (Humira™), CDP-571, CDP-870, afelimomab, lenercept, etc.), antirheumatic drugs (e.g., methotrexate, auranofin, aurothioglucose, azathioprine, sodium aurothiomalate, hydroxychloroquine sulfate, leflunomide, sulfasalazine), muscle relaxants, narcotics, non-steroidal anti-inflammatory drugsdrugs, NSAIDs), analgesics, anesthetics, sedatives, local anesthetics, neuromuscular blockers, antibacterials (e.g., aminoglycosides, antifungals, antiparasitics, antivirals, carbapenams, cephalosporins, fluoroquinolones, macrolides, penicillins, sulfonamides, tetracyclines, other antibacterials), psoriasis medications, corticosteroids, anabolic steroids, diabetes medications, minerals, nutritional drugs, thyroid medications, vitamins, calcium-related hormones, antidiarrheals, antitussives, antiemetics, antineoplastics, laxatives, anticoagulants, erythropoietin (e.g., epoetin alfa), filgrastim (e.g., G-CSF, Neupogen), sargramostim (GM-CSF, Leukine), The method further comprises administering, before, simultaneously, and / or after at least one selected from an immunological agent, an immunoglobulin, an immunosuppressant (e.g., basiliximab, cyclosporine, daclizumab), a growth hormone, a hormone replacement drug, an estrogen receptor modulator, a mydriatic, a cycloplegic, an alkylating agent, an antimetabolite, a mitotic inhibitor, a radiopharmaceutical, an antidepressant, an antimanic, an antipsychotic, an anxiolytic, a hypnotic, a sympathomimetic, a stimulant, donepezil, tacrine, an asthma medication, a beta agonist, an inhaled steroid, a leukotriene inhibitor, a methylxanthine, a cromolyn, an epinephrine or an analogue, dornase alfa (Pulmozyme), a cytokine, or a cytokine antagonist. Suitable dosages are well known in the art. See, for example, Wells et al., eds., Pharmacotherapy Handbook, 2 nd Edition, Appleton and Lange, Stamford, CT (2000), PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, CA (2000), Nursing 2001 Handbook of Drugs, 21 stedition, Springhouse Corp., Springhouse, PA, 2001; Health Professional's Drug Guide 2001, ed., Shannon, Wilson, Stang, Prentice-Hall, Inc., Upper Saddle River, NJ, each of which is incorporated herein by reference in its entirety.

[0156] Treatment Typically, the treatment of psoriasis is effected by administering an effective amount or dose of an anti-IL-23 antibody composition, on average, in the range of at least about 0.01 to 500 milligrams of anti-IL-23 antibody per kg of patient body weight per administration, and preferably at least about 0.1 to 100 milligrams of antibody per kg of patient body weight per single or multiple administrations, depending on the specific activity of the active agent contained in the composition. Alternatively, an effective serum concentration may include a serum concentration of 0.1 to 5000 μg / mL per single or multiple administrations. μ Suitable dosages are known to medical practitioners and will of course depend on the specific disease state, the specific activity of the composition administered, and the specific patient undergoing treatment. In some cases, it may be necessary to provide repeated administrations, i.e., repeated individual administrations of a specific monitored amount or quantity, to obtain the desired therapeutic dose, where the individual administrations are repeated until the desired daily dose or effect is obtained.

[0157] Preferred doses are optionally 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 7 0, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 and / or 100-500 mg / kg / dose, or any range, value or fraction thereof, or 0.1, 0.5, 0.9, 1.0, 1.1, 1.2, 1.5, 1.9, 2.0, 2.5, 2.9, 3.0, 3.5, 3.9, 4.0, 4.5, 4.9, 5.0, 5.5, 5.9, 6.0, 6.5, 6.9, 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 20, 12.5, 12.9, 13.0, 13.5, 13.9, 14.0, 14.5, 4.9, 5.0, 5.5, 5.9, 6.0, 6.5, 6.9 , 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 12, 12.5, 12.9, 13.0, 13.5, 13.9, 14, 14.5, 15, 15.5, 15.9, 16, 16.5, 16.9 , 17, 17.5, 17.9, 18, 18.5, 18.9, 19, 19.5, 19.9, 20, 20.5, 20.9, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 96, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, and / or 5000 μg / mL, or any range, value or fraction thereof.

[0158] Alternatively, the dose administered may vary depending on known factors such as the pharmacodynamic characteristics of the particular agent and its method and route of administration, the age, health and weight of the recipient, the nature and extent of the condition, type of concurrent treatment, frequency of treatment, and the desired effect. The dosage of active ingredient may typically be about 0.1 to 100 milligrams per kilogram of body weight. Typically, 0.1 to 50, preferably 0.1 to 10 milligrams per kilogram per administration, or in sustained release form, is effective to obtain the desired results.

[0159] As a non-limiting example, treatment of humans or animals can be achieved using a single dose, infusion, or multiple doses, with administration of at least one of the following: On at least one of the following days or in addition thereto, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 0.1 to 100 mg / kg per day, for example 0.5, 0.9, 1.0, 2.0, 2.5, 3.0, 3.5, 4.9, 4.9, 5.0, 5.1, 5.2, 5.3, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0, 13.0, 14.0, 15.0, 16.0, 17.0, 18.0, 19.0, 20.0, 21.0, 22.0, 23.0, 24.0, 25.0, 26.0, 27.0, 28.0, 29.0, 30.0, 31.0, 32.0, 33.0, 34.0, 35.0, 36.0, 37.0, 38.0, 39.0, 40.0, 41.0, 42.0, 43.0, 44.0, 45.0, 46.0, 47.0, 48.0, 49.0, 50.0, 51.0, 52.0, 53.0, 54.0, 55.0, 56.0, 57.0, 58.0, 59.0, 60.0, 61.0, 62.0, 63.0, 64.0, 65.0, 66.0, 67.0, 68.0, 70.0, 71.0, 72.0, 73.0, 74.0, 75.0, 76.0, 77.0, 78.0, , 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90, or 100 mg / kg of at least one antibody of the invention, either once or periodically.

[0160] Dosage forms (compositions) suitable for internal administration generally contain about 0.001 milligrams to about 500 milligrams of active ingredient per unit or container. In these pharmaceutical compositions, the active ingredient is usually present in an amount of about 0.5 to 99.999% by weight based on the total weight of the composition.

[0161] For parenteral administration, the antibody may be formulated as a solution, suspension, emulsion, particle, powder, or lyophilized powder, provided together with or separately from a pharma- ceutically acceptable parenteral vehicle. Examples of such vehicles are water, saline, Ringer's solution, dextrose solution, and 1-10% human serum albumin. Non-aqueous vehicles such as liposomes and fixed oils can also be used. The vehicle or lyophilized powder may contain additives that maintain isotonicity and chemical stability (e.g., sodium chloride, mannitol for isotonicity; buffers and preservatives for chemical stability). The formulation is sterilized by known or suitable techniques.

[0162] Suitable pharmaceutical carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, A. Osol, a standard reference text in this field.

[0163] Alternative Administration Many known and developed modes can be used in accordance with the present invention to administer a pharma- ceutically effective amount of an anti-IL-23 antibody. Although pulmonary administration is used in the following description, other modes of administration may be used in accordance with the present invention to obtain suitable results. The IL-23 specific antibodies of the present invention can be delivered in a carrier, as a solution, emulsion, colloid or suspension, or as a dry powder, using any of a variety of devices and methods suitable for administration by inhalation or by other methods described herein or known in the art.

[0164] Parenteral Formulation and Administration Formulations for parenteral administration may contain sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, hydrogenated naphthalenes, etc. as common excipients. Aqueous or oily suspensions for injection can be prepared by using appropriate emulsifiers or wetting agents and suspending agents according to known methods. Injections may be non-toxic parenterally administrable diluents such as aqueous solutions, sterile injections, or suspensions in solvents. Usable vehicles or solvents include water, Ringer's solution, isotonic saline, etc., and sterile fixed oils can be used as common solvents or suspension solvents. For these purposes, any kind of fixed oils and fatty acids, including natural, synthetic, or semi-synthetic fatty oils or fatty acids, natural, synthetic, or semi-synthetic monoglycerides, diglycerides, or triglycerides, can be used. Parenteral administration is known in the art and includes, but is not limited to, conventional injection means, gas pressurized needleless injection devices such as those described in U.S. Pat. No. 5,851,198, and laser perforator devices such as those described in U.S. Pat. No. 5,839,446, which are incorporated herein by reference in their entireties.

[0165] alternative delivery The invention further relates to administration of anti-IL-23 antibodies by parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intrabronchial, intraperitoneal, intracapsular, intrachondral, intrasinus, intracavity, intracerebellar, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, intralesional, bolus, intravaginal, rectal, buccal, sublingual, intranasal, or transdermal means. The anti-IL-23 antibody compositions may be administered parenterally (subcutaneously, intramuscularly or intravenously) or any other way, particularly for use in the form of liquid solutions or suspensions, particularly in semi-solid forms such as, but not limited to, creams and suppositories, for use in vaginal or rectal administration, in forms such as, but not limited to, tablets or capsules, for buccal or sublingual administration, or intranasally, in forms such as, but not limited to, powders, nasal drops or aerosols, or certain drugs, or with chemical enhancers such as dimethylsulfoxide to either modify the skin structure or increase the drug concentration in transdermal patches (Junginger, et al. In "Drug Permeation Enhancement;" Hsieh, DS, Eds., pp. 59-90 (Marcel Dekker, Inc. New York 1994, incorporated herein by reference in its entirety), or application of protein and peptide containing formulations to the skin (WO 98 / 53847), or application of an electric field to create a transient transport pathway, such as electroporation, or to increase the mobility of a charged drug through the skin, such as iontophoresis, or application of ultrasound, such as sonophoresis (U.S. Pat. Nos. 4,309,989 and 4,767,402), transdermally in the form of, but not limited to, a gel, ointment, lotion, suspension, or patch delivery system, using oxidizing agents that allow for this (the above publications and patents are incorporated herein by reference in their entirety).

[0166] Having generally described the invention, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended to be limiting. Further details of the invention are illustrated by the following non-limiting examples. The disclosures of all citations herein are expressly incorporated herein by reference.

[0167] Example 1: Comparison of efficacy and safety of guselkumab (GUS) with the anti-TNFα antibody adalimumab (ADA) and placebo (PBO) in patients treated over one year.

[0168] To confirm the findings from previous studies, two pivotal Phase III clinical trials were conducted: VOYAGE 1 and VOYAGE 2. Efficacy, safety, and patient-reported outcomes (PRO) findings from VOYAGE 1 were reported, comparing guselkumab with adalimumab, a widely used TNF-α inhibitor, and placebo in patients with psoriasis who were treated continuously for one year. In addition, an additional clinical trial known as VOYAGE 2 (described in Example 2) included a randomized treatment withdrawal period.

[0169] Summary of Materials / Methods for VOYAGE 1: VOYAGE 1 is a phase 3, randomized, double-blind, placebo- and active-controlled clinical trial. Eligible patients (age ≥ 18 years) had plaque psoriasis for ≥ 6 months, an Investigator's Global Assessment [IGA] score of ≥ 3, a Psoriasis Area and Severity Index [PASI] score of ≥ 12, and ≥ 10% body surface area involvement, and were candidates for systemic treatment or phototherapy. At baseline, 837 patients were randomized to receive either PBO at weeks 0 / 4 / 12, switched to GUS 100 mg at weeks 16 / 20, q8wk through 44 weeks (n=174), GUS 100 mg at weeks 0 / 4 / 12, q8wk through 44 weeks (n=329), or ADA 80 mg at week 0, 40 mg at week 1, then 40 mg q2wk through 47 weeks (n=334). The coprimary endpoints were the proportion of GUS patients versus PBO patients achieving cleared / minimal disease (IGA 0 / 1) and 90% improvement in PASI score (PASI90) at week 16. Other endpoints included IGA0 / 1, IGA0, PASI90, and PASI100 at weeks 16 / 24 / 48, and the proportion of GUS patients versus ADA patients achieving a Skin Status Questionnaire score of 0 / 1 (DLQI0 / 1), indicating no impact of infection on health-related quality of life, at weeks 24 / 48. Safety was monitored over 48 weeks.

[0170] Summary of VOYAGE 1 results: At week 16, a significantly higher (p<0.001) proportion of patients in the GUS group versus those in the PBO group achieved IGA 0 / 1 (85.1% vs. 6.9%) and PASI 90 (73.3% vs. 2.9%). GUS was also superior to ADA (p<0.001) based on the proportion of patients achieving IGA 0 / 1 (85.1% vs. 65.9%) and PASI 90 (73.3% vs. 49.7%) at week 16. Similarly, a significantly higher (p<0.001) proportion of patients achieved a response to GUS versus ADA at week 24: IGA0 (52.6% vs. 29.3%), IGA0 / 1 (84.2% vs. 61.7%), PASI100 (44.4% vs. 24.9%), and PASI90 (80.2% vs. 53.0%). The corresponding response rates at week 48 were IGA0 (50.5% vs. 25.7%), IGA0 / 1 (80.5% vs. 55.4%), PASI100 (47.4% vs. 23.4%), and PASI90 (76.3% vs. 47.9%), all p<0.001. The proportion of patients with a DLQI score of 0 / 1 among patients in the GUS vs. ADA groups was 60.9% vs. 39.5% at week 24 and 62.5% vs. 38.9% at week 48 (both p<0.001). At 48 weeks, adverse events occurred in 73.9% and 74.5% of GUS and ADA patients, respectively, and the incidence of serious adverse events was similar in the GUS and ADA groups (4.9% vs. 4.5%). Serious infections occurred in two GUS and three ADA patients. Two malignancies (prostate and breast) occurred in the GUS group. One myocardial infarction occurred in each active treatment group.

[0171] Summary conclusions of VOYAGE 1: GUS was superior to ADA in treating moderate-to-severe psoriasis and was well tolerated over 1 year of treatment.

[0172] 1444 Background: Guselkumab, an interleukin-23 (IL-23) blocker, was superior to adalimumab in treating moderate-to-severe psoriasis in a phase II clinical trial.

[0173] Objective: To compare the efficacy and safety of guselkumab with adalimumab and placebo in patients with psoriasis treated over one year.

[0174] METHODS: Patients were randomized to receive guselkumab 100 mg on weeks 0 / 4 / 12 then q8wk (n=329), placebo on weeks 0 / 4 / 16 then switched to guselkumab 100 mg on weeks 16 / 20 then q8wk (n=174), or adalimumab 80 mg on week 0, 40 mg on week 1 then 40 mg q2wk (n=334). Physician-reported outcomes (Investigator Global Assessment [IGA] score, Psoriasis Area and Severity Index [PASI]), patient-reported outcomes (PROs, Skin Status Questionnaire [DLQI], Psoriasis Symptoms and Signs Diary [PSSD]), and safety were assessed over 48 weeks.

[0175] RESULTS: Guselkumab was superior to placebo at week 16 (p<0.001) (85.1% vs. 6.9% [IGA0 / 1] and 73.3% vs. 2.9% [PASI90]). Guselkumab was also superior to adalimumab at weeks 16 (85.1% vs. 65.9%, 73.3% vs. 49.7%), 24 (84.2% vs. 61.7%, 80.2% vs. 53.0%), and 48 (80.5% vs. 55.4%, 76.3% vs. 47.9%) for IGA0 / 1 and PASI90 (p<0.001). Furthermore, guselkumab significantly improved PROs over 48 weeks. Adverse event rates were similar between treatments over 48 weeks.

[0176] LIMITATIONS: Analysis was limited to 48 weeks.

[0177] Conclusions: Guselkumab demonstrated superior efficacy compared with adalimumab and was well tolerated over 1 year in patients with psoriasis.

[0178] Materials and Methods patient The clinical trial enrolled patients aged 18 years or older with moderate to severe plaque psoriasis (i.e., Investigator's Global Assessment [IGA] ≥ 3, Psoriasis Area and Severity Index [PASI] ≥ 12, and ≥ 10% body surface area [BSA] involvement) for at least 6 months who were candidates for systemic or phototherapy. Patients were ineligible if they had a history or current symptoms of a severe, progressive, or uncontrolled medical condition, or had a current or history of malignancy, except for nonmelanoma skin cancer (NMSC), within the past 5 years. Patients with a history or symptoms of active tuberculosis (TB) were excluded. Patients were not able to participate if they had previously received guselkumab or adalimumab, other anti-TNF-α therapy within 3 months, other IL-12 / 23, IL-17, or IL-23 targeted therapy within 6 months, or any systemic immunosuppressant (e.g., methotrexate) or phototherapy within 4 weeks.

[0179] Experimental design VOYAGE 1 was a phase III, randomized, double-blind, placebo- and active-controlled clinical trial conducted at 104 sites worldwide (December 2014–April 2016). The study included an active-controlled period (weeks 0–48) comparing guselkumab to adalimumab, and a placebo-controlled period (weeks 0–16), after which placebo patients were crossed over to receive guselkumab for 48 weeks. Patients were randomized in a 2:1:2 ratio to receive guselkumab 100 mg at baseline at weeks 0, 4, and 12, then every 8 weeks for 44 weeks; placebo at weeks 0, 4, and 12, followed by guselkumab 100 mg at weeks 16 and 20, then every 8 weeks for 44 weeks; or adalimumab 80 mg at week 0, 40 mg at week 1, then 40 mg every 2 weeks for 47 weeks. Matching placebos were used to maintain blinding. Institutional review boards or ethics committees approved the protocol at participating sites, and patients provided written informed consent before study initiation.

[0180] evaluation Efficacy was assessed using the IGA, PASI, scalp-specific IGA (ss-IGA), fingernail physician global assessment (f-PGA), nail psoriasis area and severity index (NAPSI), and hand / foot PGA (hf-PGA). Patient-reported outcomes were assessed using the DLQI and the Psoriasis Symptoms and Signs Diary (PSSD). Safety monitoring included collection of adverse events (AEs) and laboratory tests.

[0181] A sensitive and drug-tolerant electrochemiluminescence immunoassay was used to detect antibodies to guselkumab, with a sensitivity of 3.1 ng / mL in guselkumab-free serum and 15 ng / mL in serum with guselkumab concentrations up to 3.125 μg / mL, exceeding the mean trough serum guselkumab level.

[0182] Statistical analysis: The co-primary endpoints were the proportion of patients in the guselkumab group achieving an IGA score of clearance or minimal disease (IGA 0 / 1) at week 16 and a 90% improvement in PASI response (PASI90) compared with placebo. Key secondary endpoints were also measured. All randomized patients were included in the primary and selected secondary efficacy analyses, and data were analyzed by randomized treatment group. The primary and key secondary analyses were tested with fixed sequence design to control for multiplicity.

[0183] The composite primary outcome and binary secondary outcomes were analyzed using the Cochran-Mantel-Haenszel (CMH) chi-square test stratified by pooled investigator site. With a sample size of approximately 750 patients, the statistical power to detect significant differences was >99% for both composite outcomes. Analysis of variance models were used to compare continuous response parameters with investigator site pooled as a covariate. All statistical tests were performed two-sided (α=0.05).

[0184] Patients who discontinued the study due to lack of efficacy or worsening psoriasis AEs, or who initiated a protocol-prohibited psoriasis treatment, were considered non-responders for binary endpoints and had baseline values ​​carried over for continuous endpoints. Other patients with missing values ​​were considered non-responders for binary endpoints (non-responder imputation method) and imputed with the last observed value for continuous endpoints (and all PSSD endpoints).

[0185] Safety analyses included all patients who received at least one dose of study drug and were summarized by actual treatment. The proportion of patients with antibodies to guselkumab was summarized among those who received at least one dose of a biologic agent.

[0186] result At baseline, 837 patients were randomized to placebo (n=174), guselkumab (n=329), or adalimumab (n=334). Overall, at 48 weeks, 6.9%, 8.5%, and 15.6% of patients in the placebo, guselkumab, and adalimumab groups, respectively, discontinued treatment. Demographic and disease characteristics were comparable across treatment groups at baseline.

[0187] Clinical response Guselkumab was superior to both placebo and / or adalimumab for the composite primary endpoint and all key secondary endpoints (all p<0.001). Compared with placebo, a significantly higher proportion of patients in the guselkumab group achieved IGA 0 / 1 (6.9% vs. 85.1%) and PASI 90 (2.9% vs. 73.3%) at week 16. In addition, the proportion achieving at least a 75% improvement in PASI (PASI 75) and IGA 0 and PASI 100 was significantly higher with guselkumab vs. placebo at week 16. Guselkumab was superior to adalimumab as measured by the proportion of patients achieving IGA 0 / 1 (85.1% vs. 65.9%), PASI 90 (73.3% vs. 49.7%), and PASI 75 (91.2% vs. 73.1%) at week 16. The significantly better response to guselkumab compared with adalimumab was maintained at week 24 (IGA0 [52.6% vs. 29.3%], IGA0 / 1 [84.2% vs. 61.7%], and PASI90 [80.2% vs. 53.0%]) and week 48 (50.5% vs. 25.7%, 80.5% vs. 55.4%, and 76.3% vs. 47.9%, respectively). In addition, a higher proportion of patients receiving guselkumab achieved a response with a higher PASI classification compared with adalimumab at week 48. After initiation of guselkumab at week 16, patients in the placebo crossover group achieved responses similar to those observed in the guselkumab group.

[0188] Topical Psoriasis Scale Localized psoriasis was evaluated based on ss-IGA, f-PGA, NAPSI, and hf-PGA assessments. The proportion of patients achieving ss-IGA 0 / 1 (absent / very mild scalp psoriasis) in the guselkumab group was significantly higher compared to placebo at week 16 (83.4% vs. 14.5%, p<0.001), and a significantly better response to guselkumab versus adalimumab was observed at weeks 24 (p<0.001) and 48 (p=0.045). The proportion of patients achieving f-PGA 0 / 1 (eliminated / minimal) and improvement rate in NAPSI were significantly higher with guselkumab versus placebo at week 16 (p<0.001). f-PGA responses were comparable at week 24, but guselkumab was superior to adalimumab at week 48 (p=0.038). The mean improvement in NAPSI with guselkumab was significantly higher than placebo at week 16 (p<0.001) and was similar between guselkumab and adalimumab at weeks 24 and 48. Finally, the proportion of patients achieving hf-PGA 0 / 1 (cleared / almost cleared) was significantly higher with guselkumab versus placebo at week 16, and the response to guselkumab was superior to adalimumab at weeks 24 and 48 (p<0.001).

[0189] Health-related quality of life measures At week 16, improvement from baseline in DLQI was significantly greater in the guselkumab group compared with placebo (mean change, -0.6 vs. -11.2), as was the proportion of patients achieving DLQI 0 / 1 (no effect of psoriasis on HRQoL) (both p<0.001). At weeks 24 and 48, improvement from baseline in DLQI and the proportion of patients achieving DLQI 0 / 1 were significantly greater with guselkumab compared with adalimumab (p<0.001).

[0190] At week 16, the improvement from baseline in PSSD symptom scores was significantly higher with guselkumab versus placebo (mean change, -3.0 vs. -41.9), and the mean change in PSSD sign scores was similarly favorable for guselkumab (both p<0.001). Similarly, at weeks 24 and 48, the mean change in PSSD symptom and sign scores in the guselkumab group was significantly greater than that in the adalimumab group (p<0.001). The proportion of patients achieving a PSSD symptom score=0 with guselkumab and adalimumab, respectively, was 36.3% and 21.6% at week 24, and the significantly better response to guselkumab was sustained at week 48 (p<0.001). Similar results were observed for the proportion of patients achieving a PSSD sign score=0 at weeks 24 and 48 (p<0.001).

[0191] Safety outcomes During the placebo-controlled period (weeks 0–16), the proportion of patients with at least one AE was comparable across treatment groups, with the most commonly reported events being nasopharyngitis and upper respiratory tract infection in all three groups. Serious AEs (SAEs) and AEs leading to discontinuation of study drug occurred less frequently and in similar proportions of patients for each treatment. The incidence of infections overall and infections requiring antibiotic treatment was comparable across treatment groups. Two patients in the adalimumab group experienced serious infections (both cellulitis). One case of NMSC (i.e., basal cell carcinoma [BCC]) was reported in the guselkumab group, and no other malignancies occurred in either group. One myocardial infarction (i.e., major adverse cardiovascular event [MACE]) occurred in each active treatment group through week 16.

[0192] The types and patterns of AEs reported over the 48 weeks were similar to those reported during the placebo-controlled period. The proportions of patients with at least one AE, AE leading to discontinuation, or SAE were similar in the guselkumab and adalimumab groups. Between weeks 16 and 48, serious infections were reported in two patients in the guselkumab group (i.e., one thigh abscess with cellulitis and the other postoperative wound infection) and two in the adalimumab group (i.e., one abdominal abscess and the other staphylococcal pneumonia leading to a fatal outcome in a patient with abdominal wall cellulitis reported earlier). Overall infections and infections requiring antibiotic treatment occurred at similar rates across treatment groups. No AEs of active tuberculosis or opportunistic infections were reported during the study. Two additional NMSC (i.e., one BCC in each of the guselkumab and adalimumab groups) and two malignancies (i.e., prostate and breast cancer in the guselkumab group) were reported at 48 weeks. No additional MACE occurred after 16 weeks. One suicide attempt was reported in an adalimumab patient. The incidence of candidiasis and neutropenia was low and similar between the guselkumab and placebo groups (data not shown), and no Crohn's disease events were reported at 48 weeks.

[0193] Through 48 weeks, the proportion of patients with ISR (2.2% vs. 9.0%) and the proportion of injections associated with ISR (0.5% vs. 1.2%) were lower with guselkumab compared with adalimumab, with most ISRs considered to be mild. The incidence of laboratory abnormalities was low and did not differ between groups (data not shown). Antibodies to guselkumab were detected in 26 / 492 patients (5.3%) through 44 weeks, with titers generally low (81% ≦1:320). No clear association was observed between the occurrence of antibodies and either reduced efficacy or the occurrence of ISRs (data not shown).

[0194] Consideration The findings in the VOYAGE 1 trial, together with the results of VOYAGE 2 described in Example 2 below, confirmed that two injections of 100 mg of guselkumab (at weeks 0 and 4) and maintenance therapy every 8 weeks effectively treated moderate to severe infections. Guselkumab was superior to placebo by a substantial margin at week 16 using two rigorous endpoints (IGA0 / 1 and PASI90). The onset of action of guselkumab was rapid, with evidence of significant response as early as week 2 compared to placebo. Guselkumab was also superior to adalimumab, a widely used and highly effective subcutaneous TNF-α inhibitor, in the week 16 endpoints of IGA0 / 1, PASI90, and PASI75. Response rates continued to improve beyond week 16 with guselkumab. At weeks 24 and 48, approximately half of all patients in the guselkumab group achieved complete clearance (IGA0), which is associated with optimal HRQoL for patients with psoriasis. Patient-reported outcome measures (PSSD and DLQI) based on total change or minimal / no impact on HRQoL, or absence of symptoms or signs of psoriasis demonstrated a response of guselkumab superior to placebo at week 16 and superior to adalimumab at weeks 24 and 48.

[0195] This study evaluated multiple sites on the body where psoriasis is difficult to treat, and guselkumab was highly effective in all sites based on rigorous endpoints. Guselkumab was superior to placebo at week 16 and to adalimumab at weeks 24 / 48, demonstrating complete or near complete clearance of scalp and hand / foot psoriasis in at least 75% of guselkumab-treated patients. Based on both the proportion of patients with cleared / minimal fingernail psoriasis (f-PGA 0 / 1) and the mean improvement rate in NAPSI, guselkumab was superior to placebo at week 16. Nail responses were comparable to active treatment at weeks 24 and 48, and guselkumab was superior to adalimumab for f-PGA 0 / 1 at week 48 (75% vs. 62%).

[0196] The incidence and type of AEs, SAEs, and laboratory abnormalities were similar between the guselkumab and placebo groups through 16 weeks, and between the guselkumab and adalimumab groups through 48 weeks. The incidence of serious infections, malignancies, and MACEs was low across treatment groups. No notable differences in the incidence of neutropenia or candidiasis were observed between the guselkumab and control groups. No Crohn's disease AEs occurred in either treatment group, and one suicide attempt was reported in the adalimumab group. The number and percentage of injections of patients with ISR were higher with adalimumab compared to guselkumab. The size and duration of this study may not allow for evaluation of rare events or those with long latency periods, but longer-term treatment will be evaluated in the ongoing extension of the study.

[0197] VOYAGE1 confirms the role of IL-23 in the pathogenesis of psoriasis. TNF-α inhibitors are effective in many diseases, and TNF-α is involved in normal systemic inflammation and immunological processes. Selective targeting of the IL-23 pathway provides more specific cytokine inhibition for psoriasis with a higher degree of efficacy while maintaining a good safety profile when compared to TNF-α blockers. IL-23 is a key driver of Th17 cell differentiation and survival, and an upstream regulator of IL-17A, a central inflammatory effector cytokine involved in the pathogenesis of psoriasis. In addition, IL-23 stimulates the production of other Th17 cytokines (e.g., IL-22) by other cell types, including innate lymphoid cells type 3 (ILC3) cells and γδ T cells. Thus, inhibition of IL-23 blocks downstream production of IL-17A, IL-22, and other cell types. Because many IL-17A-producing cells depend on IL-23 for survival, inhibition of IL-23 may reduce the numbers of these pathogenic cells, which may explain the long duration of effect and allow for a convenient dosing interval for guselkumab compared to both anti-TNF-α and anti-IL-17 agents.

[0198] This finding, together with those from VOYAGE 2, demonstrates superior efficacy of guselkumab compared with adalimumab in psoriasis, efficacy in localized disease of the scalp, nails, and hands / feet, and a clear safety profile. The favorable safety profile through one year is not unexpected, given that it reaffirms the long-term safety findings reported for a related treatment, ustekinumab (which blocks both IL-12 and IL-23). ​​Extension of the study will continue to explore the efficacy and safety of guselkumab and increase understanding of long-term IL-23 blockade based on the ustekinumab studies.

[0199] Example 2: VOYAGE2 Results To confirm the therapeutic potential of guselkumab, the phase 3 VOYAGE 1 and VOYAGE 2 trials evaluated the efficacy and safety of guselkumab versus placebo and adalimumab. VOYAGE 1 evaluated continuous 1-year treatment, and VOYAGE 2 evaluated the efficacy and safety of interrupted treatment, as treatment gaps frequently occur in clinical practice. In addition, VOYAGE 2 evaluated the transition from adalimumab to guselkumab, providing clinically relevant information for patients switching biologics.

[0200] Materials and Methods patient Adults (age ≥18 years) with moderate to severe plaque psoriasis were eligible. Key inclusion / exclusion criteria are summarized in VOYAGE1 above. The study protocol was approved by the Institutional Review Board at each site, and written informed consent was provided by all patients.

[0201] Experimental design VOYAGE2 was a phase 3, multicenter, randomized, double-blind, placebo- and adalimumab-controlled trial (NCT02207244) conducted at 115 centers worldwide between November 2014 and June 2016. The study consisted of a placebo-controlled period (weeks 0-16), an active-controlled period (weeks 0-28), and a randomized withdrawal and retreatment period (weeks 28-72). Herein, we present study results through 48 weeks. Patients were randomized 2:1:1 at baseline to receive guselkumab 100 mg at weeks 0, 4, 12, and 20; placebo at weeks 0, 4, and 12 followed by guselkumab at weeks 16 and 20; or adalimumab 80 mg at week 0 and 40 mg at week 1, then every 2 weeks (q2w) for 23 weeks.

[0202] At week 28, guselkumab-treated patients who achieved 90% from baseline in the Psoriasis Area and Severity Index (PASI90, responders) were re-randomized in a 1:1 ratio to guselkumab or placebo. At ≥50% loss in PASI response at week 28, patients were re-treated with guselkumab followed by another dose 4 weeks later, then q8w thereafter. Guselkumab non-responders continued guselkumab treatment. Placebo → week 28 guselkumab non-responders continued guselkumab q8w, whereas responders received placebo q8w starting at week 28. At ≥50% loss in PASI response at week 28, patients were re-treated with guselkumab followed by another dose 4 weeks later, then q8w thereafter. Adalimumab non-responders started guselkumab at week 28, received another dose 4 weeks later, then q8w thereafter. Adalimumab responders received placebo and, upon ≥50% loss in PASI response at week 28, started guselkumab, received another dose 4 weeks later, then q8w thereafter. Both guselkumab and adalimumab placebo were administered as needed to maintain blinding.

[0203] Efficacy and safety evaluation Efficacy was assessed through 24 weeks, and safety was assessed through 28 weeks. Key efficacy outcomes, including generalized and localized psoriasis, patient-reported outcomes, and safety assessments, are detailed in VOYAGE 1. In addition, the Medical Outcomes Study 36-item Short Form (SF-36) was evaluated in this study.

[0204] Study Evaluation Items The co-primary endpoints were the proportion of patients achieving an IGA score of eliminated (0) or minimal (1) at week 16 and the proportion of patients achieving a PASI 90 response at week 16 in the guselkumab vs placebo groups. Key secondary endpoints were also measured. Localized psoriasis endpoints evaluating the scalp, fingernails, and hands / feet are described in detail elsewhere.

[0205] statistical analysis All randomized patients were included in the primary and several secondary analyses according to their assigned treatment group. To evaluate the main secondary endpoint of maintenance dose on withdrawal from treatment, all patients who underwent a second randomization and had a PASI assessment after 28 weeks were included in the analysis.

[0206] The composite primary outcome and binary secondary outcomes were analyzed using the Cochran-Mantel-Haenszel (CMH) chi-square test stratified by investigator site (α = 0.05). Continuous response parameters were compared with site as a covariate using analysis of variance models. For time to loss of PASI90 response, the log-rank test stratified by site was used.

[0207] Patients who discontinued study treatment due to lack of efficacy or AEs of worsening psoriasis, or who initiated a protocol-prohibited drug / therapy that could improve psoriasis, were considered treatment failures. Patients who met treatment failure criteria before week 16 and those who did not return for the week 16 assessment were considered non-responders to the week 16 primary endpoint. For patients randomized to placebo, only patients crossing over to receive guselkumab at / after week 16 were included in the post-week 16 efficacy analysis.

[0208] To control for the overall type I error rate, the primary and secondary analyses were tested in a fixed order, with the first primary secondary endpoint being tested only if the composite primary endpoint was met, and subsequent endpoint(s) being tested only if the preceding endpoint in the sequence was met.

[0209] Safety analyses included all patients who received at least one dose of study drug. Adverse events and serious adverse events (SAEs) were grouped according to treatment received. Antibodies to guselkumab were analyzed.

[0210] result Patient demographics and baseline demographic characteristics A total of 1279 patients were screened, and 992 were randomly assigned 2:1:1 to receive guselkumab (n=496), placebo (n=248), or adalimumab (n=248) (Figure 2). Overall, 9.7% (96 / 992) of patients discontinued study drug through week 48 (guselkumab: 7.9%, placebo: 11.7%, adalimumab: 11.3%). Baseline demographic and disease characteristics were generally comparable between groups.

[0211] Effectiveness Guselkumab was superior to both placebo and / or adalimumab for the co-primary endpoint and all primary secondary endpoints (all p<0.001).

[0212] Placebo-controlled period (weeks 0-16) At week 16, a significantly greater proportion of guselkumab patients achieved an eliminated (0) or minimal (1) IGA compared with placebo patients (84.1% vs. 8.5%, p<0.001) and a PASI 90 response (70.0% vs. 2.4%, p<0.001) (composite primary endpoint). Significantly higher PASI improvement rates were observed as early as week 2 with guselkumab versus placebo patients (p<0.001). In addition, at week 16, a significantly higher proportion of guselkumab patients achieved PASI 75 and PASI 100 responses compared with placebo patients. Guselkumab patients achieved greater improvements in all localized psoriasis outcome measures, including ss-IGA, f-PGA, NAPSI, and hf-PGA, compared with placebo at week 16. As in VOYAGE 1, guselkumab was superior to placebo at week 16 in all patient-reported outcomes, including the Skin Status Questionnaire (DLQI) and Diary of Psoriasis Symptoms and Signs (PSSD), in addition to the SF-36.

[0213] Comparison period (0-24 weeks) A significantly greater proportion of patients in the guselkumab group achieved IGA0 / 1, PASI90, and PASI75 responses at week 16. At week 24, significantly higher response rates were maintained for IGA0 (51.5% vs. 31.5%), IGA0 / 1 (83.5% vs. 64.9%), PASI90 (75.2% vs. 54.8%), and PASI100 (44.2% vs. 26.6%) in guselkumab versus adalimumab patients. Consistent with VOYAGE 1, greater improvements in localized psoriasis disease scores were observed at week 24 in guselkumab patients compared to adalimumab patients, except for improvement rates in f-PGA0 / 1 and NAPSI, which were comparable. At week 24, the proportion of patients achieving a DLQI of 0 / 1, mean change in PSSD symptom and sign scores, a PSSD symptom score of 0, and a sign score of 0 were significantly greater in the guselkumab group than in the adalimumab group (p<0.001).

[0214] Randomized withdrawal and retreatment period (28-48 weeks) PASI90 responses were better maintained in guselkumab responders at 28 weeks who continued guselkumab (maintenance group) versus responders re-randomized to placebo (withdrawal group). The median time to loss of PASI90 response was 15.2 weeks in patients randomized to the withdrawal group. Among patients who withdrew from guselkumab at week 28, PASI90 response rates began to diverge from the maintenance group at week 32. Through 48 weeks, 88.6% of maintenance patients maintained a PASI90 response versus 36.8% of withdrawal patients. In addition, at week 48, clinical responses (IGA, PASI) were significantly higher in the maintenance group than in the withdrawal group (p<0.001). Improvements from baseline in DLQI and PSSD symptom or sign scores were also significantly greater at week 48 in the maintenance group versus the withdrawal group (both p<0.001). Through 48 weeks, a small number of patients (n=16) were retreated with guselkumab.

[0215] Switching to guselkumab in adalimumab non-responders Overall, 112 adalimumab non-responders initiated guselkumab at week 28 (5 weeks after the last adalimumab dose). In these patients, PASI90 (vs. baseline) and PASI100 response rates increased after switching, reaching 66.1% and 28.6%, respectively, at week 48.

[0216] safety Placebo-controlled period (weeks 0-16) The proportion of patients with at least one AE, AEs leading to discontinuation, and SAEs were comparable between the guselkumab and placebo groups. The most commonly reported events were nasopharyngitis, headache, and upper respiratory tract infection. The incidence of infections, infections requiring treatment, and serious infections were similar between groups. No malignancies or nonmelanoma skin cancers (NMSCs) were reported through week 16. One major adverse cardiovascular event (MACE) (myocardial infarction [MI]) occurred in the adalimumab group. Compared with guselkumab patients, a higher proportion of adalimumab patients had injection site reactions (ISRs) (6.9% vs. 2.6%) and injection sites resulting in ISRs (1.5% vs. 0.9%). All injection site reactions were mild in severity.

[0217] Comparison period (0-28 weeks) The types of AEs were similar to those reported during the placebo-controlled period. The proportions of patients with at least one AE, AEs leading to discontinuation, and SAEs were comparable between the guselkumab and adalimumab groups. Infections and infections requiring treatment were also comparable between the guselkumab and adalimumab groups. Three serious infections were reported in the guselkumab group (bronchitis, erythema, and soft tissue infection) and adalimumab groups (two cases of tuberculosis [one diffuse], and one injection site abscess), respectively. One malignancy (prostate cancer) and two NMSCs (one squamous cell carcinoma [SCC] in the guselkumab group and one basal cell carcinoma [BCC] in the placebo → guselkumab group) were reported. Two MACEs (one MI in each of the guselkumab and adalimumab groups, respectively) were reported.

[0218] Randomized withdrawal and retreatment period (28-48 weeks) From weeks 28 to 48, no patients discontinued due to AEs, and one serious infection (appendicitis) was reported in the maintenance group. No additional malignancies, NMSC, or MACE were reported. No AEs were reported among the 16 retreated patients.

[0219] Further safety through 48 weeks Through week 48, one additional BCC and one additional SCC of the skin occurred in the placebo → guselkumab group (data not shown). Between weeks 28 and 48, one additional MACE (MI) was reported in the placebo → guselkumab patient. No serious infections, malignancies, or MACE events occurred in the adalimumab → guselkumab group. No deaths, opportunistic infections, hypersensitivity, or anaphylactic reactions occurred through week 48. The incidence of laboratory abnormalities was low and comparable between treatment groups through week 48. Antibodies to guselkumab were detected in 57 / 869 (6.6%) patients through week 48, with titers generally low (88% ≤ 1:160). No clear association was observed between the occurrence of antibodies and either reduced efficacy or the occurrence of ISRs (data not shown).

[0220] Identification and evaluation of clinical parameters as single predictors for maintenance of PASI90 response after withdrawal of guselkumab treatment in patients with chronic plaque psoriasis At week 28, PASI 90 responders (patients with a 90% improvement in the Patient Psoriasis Area and Severity Index (PASI) score) randomized to guselkumab at baseline were re-randomized to either continue on guselkumab 100 mg or switch to placebo (group 1c) (Figure 1). Patients randomized to placebo at baseline who crossed over to guselkumab at weeks 16 and 20 and achieved a PASI 90 response at week 28 were also switched to placebo (group 2b) (Figure 1). Thirty-seven percent of patients in group 1c and 42% of patients in group 2b demonstrated long-term maintenance of skin improvement (maintained PASI 90 response at week 48, 28 weeks after the last guselkumab dose at week 20), suggesting that guselkumab can induce sustained remission in some patients.

[0221] A number of clinical parameters were evaluated for their association with long-term maintenance of PASI 90 response after withdrawal of guselkumab treatment in 182 patients in group 1c (Figure 1). Clinical parameters evaluated included disease duration, age at disease onset, body mass index (BMI), diseased body surface area (BSA), Investigator Global Assessment (IGA) score, psoriatic arthritis status, and prior use of biologics at baseline, as well as IGA score and PASI improvement at week 28. Among the various parameters evaluated, shorter disease duration (p<0.005) (Figure 2), lower BMI at baseline (p<0.001) (Figure 3), achieving IGA 0 at week 28 (p<0.001) (Figure 4), and achieving a PASI 100 response at week 28 (p<0.005) (Figure 5) were associated with a drug-free long-term response of sustained PASI 90 7 months after the last dose of guselkumab. Associations were assessed by Welch's t-test or Fisher's exact test. Single predictor models were constructed based on each of these identified parameters using univariate logistic regression and evaluated by the area under the curve (AUC) of the receiver operating characteristic curve. Group 1c was used as the training set and the model was validated in another cohort, group 2b (Figure 6, Figure 7, Figure 8, and Figure 9).

[0222] Identification and evaluation of serum protein biomarkers associated with sustained clinical benefit after withdrawal of guselkumab treatment in patients with chronic plaque psoriasis Inflammatory proteins or cytokines of the IL-23 / Th17 axis are believed to be the main drivers of psoriasis pathogenesis and are involved in the initiation and chronic stages of the disease. IL17A, IL17F, and IL22 are produced by several types of immune cells, such as innate lymphocytes and T cells, as a result of IL23 binding to the IL23 receptor on the surface of those cells. IL23 is mainly produced by antigen-presenting cells, such as macrophages and dendritic cells. IL17A, IL17F, and IL22, among other cytokines, then bind to receptors on keratinocytes, triggering the production of chemokines, which then attract more immune cells. Keratinocytes also respond to inflammatory cytokines by producing antimicrobial proteins, such as beta-defensins, and other inflammatory and stress-induced proteins, such as S100 proteins, which act as extracellular alarmins or damage-associated molecular pattern factors. Cytokines and other proteins produced in the skin during psoriasis are often found at elevated concentrations in the circulating blood of psoriasis. It is not known whether these cytokines / proteins found in serum are produced first in the skin and then enter the periphery or are produced by cells in the circulation. These elevated levels of cytokines and other inflammatory proteins are measured in the circulation and serve as biomarkers of disease activity and potentially for predicting efficacy response and maintenance of response. In VOYAGE 2, serum from peripheral blood was collected at baseline and at various time points during and after treatment with guselkumab was stopped. Various methods, including the Mesoscale method and the highly sensitive Singulex method, were used to determine the concentrations of various cytokines and other proteins in serum from patients at various time points throughout the study. Baseline levels of several cytokines were elevated at baseline, including IL17A, IL17F, IL22, IL23A, CCL22 / MDC, IL8, and CCL4 / MIP-1β at baseline. The association between the levels of these various cytokines at baseline and at various time points throughout the study was evaluated for association with maintenance of efficacy 28 weeks after the last dose of guselkumab at week 20.Lower serum levels of IL17F (p<0.03) (Figure 10) and MIP1β (p<0.02) (Figure 11) at baseline were associated with a higher likelihood of sustaining PASI90 responses. Lower levels of IL17A and IL22 were also associated with drug-free remission, but these associations did not reach statistical significance. Single predictor models were constructed based on baseline serum concentrations of IL17F or MIP1β using univariate logistic regression in group 1c as the training set and assessed by the area under the curve (AUC) of the receiver operating characteristic curve (Figure 12).

[0223] Identification and evaluation of circulating guselkumab drug levels as a pharmacokinetic marker associated with sustained clinical benefit after discontinuation of guselkumab treatment in patients with chronic plaque psoriasis In VOYAGE 2, guselkumab concentrations in patients' blood were monitored throughout the study. Higher guselkumab blood concentrations at week 28 were associated with drug-free maintenance of PASI 90 response 28 weeks after the last dose of guselkumab (Figure 13). Modeling of this parameter using multivariate regression analysis yielded AUCs of 0.77 and 0.63 in the training (group 1c) and validation (group 2b) cases, respectively (Figure 14).

[0224] Comparison of single-parameter predictor models Clinical parameters that were evaluated and found to be associated with a drug-free long-term response of maintenance of PASI 90 7 months after the last dose of guselkumab included shorter disease duration (p<0.005) (Figure 2), lower BMI at baseline (p<0.001) (Figure 3), achieving IGA0 at week 28 (p<0.001) (Figure 4), and achieving a PASI 100 response at week 28 (p<0.005) (Figure 5), which were then used to build single predictor models using univariate logistic regression (Figures 6-9, 12, and 14). Various single parameter predictor models were evaluated by ordering them based on the AUC of the ROC curve and are shown in Figure 15. A comparison of predictor models built by combining various baseline parameters including the AUC of the ROC curve is shown in Figure 16. A comparison of selected predictor models built by combining multiple parameters including baseline parameters and parameters during or after treatment with guselkumab is shown in Figure 17.

[0225] Identification and evaluation of a predictive model combining clinical parameters and pharmacokinetic markers for long-term maintenance of PASI90 response after withdrawal of guselkumab treatment in patients with chronic plaque psoriasis To improve the predictive ability of long-term maintenance of PASI90 response, models were constructed by combining clinical parameters and pharmacokinetic markers using multivariate logistic regression and evaluated by the area under the curve (AUC) of the receiver operating characteristic curve. The model combining disease duration, baseline BMI, and IGA0 response at week 28 had AUCs of 0.72 and 0.71 in the training and validation sets, respectively (Figure 18). The model combining disease duration, IGA0 response, and GUS concentration at week 28 had AUCs of 0.83 and 0.74 in the training and validation sets, respectively (Figure 19).

[0226] Methods for using a predictive model to predict maintenance of a PASI score of less than 3 by achieving a PASI 90 response at week 28 following initial administration of guselkumab at weeks 0, 4, 12, and 20 followed by guselkumab maintenance therapy at every 16-week dosing interval A logistic regression model based on disease duration (PSODUR), baseline body mass index (BMI), and achieving Investigator Global Assessment (IGA) skin clearance at week 28 (IGA0_WK28) was generated to predict maintenance of a PASI score <3 by achieving a PASI90 response at week 28, administered with maintenance guselkumab treatment at 16-week dosing intervals following initial doses of guselkumab at weeks 0, 4, 12, and 20.

[0227] The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(3.3492-0.03244 x PSODUR-0.05807 x BMI + 1.6254 x IGA0_WK28))).

[0228] The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 67% of guselkumab-treated patients had a predictive score greater than 0.72, and 90% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 16-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28.

[0229] Methods for using a predictive model to predict maintenance of a PASI score of less than 3 by achieving a PASI 90 response at week 28 following initial administration of guselkumab at weeks 0, 4, 12, and 20 followed by guselkumab maintenance therapy at every 20-week dosing interval A logistic regression model based on disease duration (PSODUR), baseline body mass index (BMI), and achieving investigator's global assessment (IGA) clearance at week 28 (IGA0_WK28) was generated to predict maintenance of a PASI score <3 by achieving a PASI90 response at week 28, administered with maintenance guselkumab therapy at 20-week dosing intervals following initial doses of guselkumab at weeks 0, 4, 12, and 20.

[0230] The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(2.6101-0.02557 x PSODUR-0.05892 x BMI + 1.0128 x IGA0_WK28))).

[0231] The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, whereas the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line indicates that approximately 33% of guselkumab-treated patients had a predictive score greater than 0.79, and 85% of these patients received guselkumab at 20-week dosing intervals after the initial administration of guselkumab at weeks 0, 4, 12, and 20; This represents an example in which a PASI score of less than 3 could be maintained by achieving a PASI90 response at 28 weeks.

[0232] A logistic regression model based on disease duration (PSODUR), achieving investigator global assessment (IGA) clearance at week 28 (IGA0_WK28), and blood guselkumab levels at week 28 (GUS_WK28) was generated to predict maintenance of a PASI score <3 by receiving guselkumab therapy at weeks 0, 4, 12, and 20 followed by administration of guselkumab maintenance therapy at 20-week dosing intervals and achieving a PASI90 response at week 28. FIG.

[0233] The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(-0.1615-0.02004 x PSODUR + 1.0145 x IGA0_WK28 + 0.8199 x GUS_WK28))).

[0234] The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 42% of guselkumab-treated patients had a predictive score greater than 0.75, and 89% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 20-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28.

[0235] Methods for using a predictive model to predict maintenance of a PASI score of less than 3 by achieving a PASI 90 response at week 28 following initial administration of guselkumab at weeks 0, 4, 12, and 20 followed by guselkumab maintenance therapy at every 24-week dosing interval A logistic regression model based on disease duration (PSODUR), achieving investigator's global assessment (IGA) clearance at week 28 (IGA0_WK28), and blood guselkumab levels at week 28 (GUS_WK28) was generated to predict maintenance of a PASI score <3 by receiving guselkumab at weeks 0, 4, 12, and 20 followed by maintenance guselkumab therapy at 24-week dosing intervals and achieving a PASI90 response at week 28. FIG.

[0236] The equation for the logistic regression model is defined as predicted score = 1 / (1 + exp(-(-1.1498-0.02462 x PSODUR + 1.2800 x IGA0_WK28 + 0.9031 x GUS_WK28))).

[0237] The probability of maintaining a PASI score below 3 among patients with a predictive score greater than a given cutoff is defined as the positive predictive value (PPV) and is shown as a solid line, while the proportion of patients with a predictive score greater than a given cutoff after receiving guselkumab at weeks 0, 4, 12, and 20 is shown as a dotted line. The dotted line represents an example where approximately 40% of guselkumab-treated patients had a predictive score greater than 0.60, and 78% of these patients were able to maintain a PASI score below 3 by receiving guselkumab at 24-week dosing intervals following initial administration of guselkumab at weeks 0, 4, 12, and 20 and achieving a PASI 90 response at week 28.

[0238] Consideration VOYAGE 2 confirms the results of VOYAGE 1, supporting that guselkumab is highly effective in treating a broad range of moderate-to-severe psoriasis populations. Guselkumab was superior to placebo in the 16-week composite primary endpoints of excluded / minimal IGA and PASI 90 response. Guselkumab was also superior to adalimumab in the 16-week endpoints of excluded / minimal IGA, PASI 75 / 90, and in excluded IGA and PASI 90 / 100 through week 24. Guselkumab also successfully treated challenging localized psoriasis, including on the scalp, nails, and hands / feet. Investigator-assessed improvements were reflected in improvements in patient-reported outcomes assessed in VOYAGE 1 (DLQI and PSSD, a newly validated scale measuring signs and symptoms of psoriasis). In addition, significant improvements in quality of life (QoL) (SF-36) were reported in VOYAGE 2. Combined robust and comprehensive results from VOYAGE 1 and VOYAGE 2 demonstrated superiority over both placebo and adalimumab.

[0239] Consistent with findings using other biologics for psoriasis, VOYAGE 2 demonstrated that maintenance therapy was superior to discontinuation therapy and that blockade of IL-23 did not permanently reverse the underlying mechanisms of psoriasis, at least in most patients. Unlike VOYAGE 1, in which patients continued treatment for 48 weeks, VOYAGE 2 randomized PASI 90 responders at week 28 to continue guselkumab or receive placebo. Guselkumab-treated patients maintained responses, including PASI 90, PASI 100, and excluded IGA, while placebo patients experienced a slow relapse of psoriasis and a decline in QoL. The median time to loss of PASI 90 response for withdrawal patients was 15.2 weeks, which is associated with a significant (often >50%) discontinuation rate with psoriasis biologics use throughout the year.

[0240] Guselkumab was also effective in treating adalimumab non-responders (those who did not achieve PASI 90). Patients were classified as responders or non-responders at week 28, 5 weeks after the last adalimumab injection. At week 20 after guselkumab treatment, 2 / 3 of 112 adalimumab non-responders who switched to guselkumab achieved PASI 90 (vs. baseline). Determining the proportion of patients with suboptimal responses who achieve treatment goals may significantly impact treatment decisions for patients seeking greater clearance as more effective psoriasis therapies continue to enter the market. The mechanism of action of guselkumab's broader efficacy is unknown, but in psoriasis, tumor necrosis factor-α (released from CD163+ macrophages / CD11c+DCs) and IL-17A (released from neutrophils, Th17, and Tc17 cells) are effector cytokines that act primarily on keratinocytes. IL-23 can be seen as a global master cytokine for psoriasis, as it promotes T cell activation and induces IL-17 production. In addition, IL-23 induces innate cells, such as Th17, Th17 cells, and innate lymphocytes, to produce IL-22, another cytokine involved in the pathogenesis of psoriasis. Therefore, targeting the IL-23 pathway with an antibody such as guselkumab can provide a more effective and durable response.

[0241] The safety profile of guselkumab in this study was consistent with VOYAGE 1. The incidence and type of AEs, SAEs, and laboratory abnormalities were comparable to placebo through 16 weeks and to adalimumab through 28 weeks. Through 48 weeks, the incidence of serious infections, malignancies, and MACEs was low across treatment groups. Overall, there were two cases of TB, both in adalimumab patients. There were five malignancies in guselkumab-treated patients, four of which were NMSC (two BCC and two SCC). ISRs occurred more frequently in adalimumab patients. While the safety profile of guselkumab was favorable, the size and duration of the study were insufficient to detect rare events.

[0242] VOYAGE 2 has several other limitations. The comparison of guselkumab with adalimumab in VOYAGE 2 was limited to 24 weeks, whereas VOYAGE 1 extended the comparison with adalimumab to 48 weeks. More importantly, at week 48, few patients in VOYAGE 2 who withdrew from active therapy had lost sufficient response to allow evaluation of the efficacy and safety of treatment. However, these numbers increase at later time points.

[0243] In conclusion, VOYAGE 2 confirms the results of VOYAGE 1, which showed that guselkumab had superior efficacy and comparable safety to adalimumab when administered in a simple drug regimen of a single 100 mg injection at weeks 0, 4, and every 8 weeks. These results suggest that guselkumab may be an important addition to psoriasis treatment options. In addition, VOYAGE 2 provides important data regarding the need for continued therapy with guselkumab to maintain the highest level of response, as well as the need for successful transition from adalimumab to guselkumab.

[0244] [Table 2]

[0245] US Regulatory Approval Guselkumab, an anti-IL-23 specific antibody, has been approved for marketing in the United States by the U.S. Food and Drug Administration through a Biologics License Application for the treatment of adult patients with moderate to severe plaque psoriasis who are candidates for systemic therapy or phototherapy. The initial approved dose is 100 mg administered by subcutaneous injection at weeks 0, 4, and every 8 weeks thereafter. The antibody is present in a concentration of 100 mg / mL in single-dose prefilled syringes. The following aspects may be included. [1] A method for predicting sustained response to IL-23 antibody therapy and treating a patient with psoriasis, the method comprising: a. administering to said patient an IL-23 antibody; b. obtaining or having obtained a biological sample from said patient; c. measuring or having measured at least one indicator from the patient or the biological sample before and during treatment with the IL-23 antibody, the indicator being selected from the group consisting of a clinical parameter, a pharmacokinetic marker, and a serum biomarker; d. generating a predictive patient profile based on the measured indicators; and e. administering said maintenance therapy of said IL-23 antibody according to said predicted patient profile. [2] The method according to [1] above, wherein the IL-23 antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO: 1, the CDRH2 amino acid sequence of SEQ ID NO: 2, and the CDRH3 amino acid sequence of SEQ ID NO: 3, and the light chain variable region comprising the complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO: 4, the CDRL2 amino acid sequence of SEQ ID NO: 5, and the CDRL3 amino acid sequence of SEQ ID NO: 6. [3] The method according to [2] above, wherein the anti-IL-23 antibody comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 7 and the light chain variable region amino acid sequence of SEQ ID NO: 8. [4] The clinical parameters of the predictive patient profile include: a. Illness period; B.BMI, c. IGA score, and The method according to any one of the above [1] to [3], wherein the anti-inflammatory agent is selected from the group consisting of d. PASI. [5] The method according to any one of the above [1] to 3], wherein the pharmacokinetic parameter is the blood concentration of the IL-23 antibody. [6] The method according to any one of [1] to [3] above, wherein the serum protein biomarkers of the predicted patient profile are selected from the group consisting of IL17F and MIP1β. [7] The method according to any one of the above-mentioned [1] to [3], wherein the IL-23 antibody is administered at an initial administration, 4 weeks after the initial administration, and every 8 weeks after the administration at the 4th week. [8] The method according to any of the above [1] to [3], wherein the patient is identified as a responder to the IL-23 antibody and having a PASI90, PASI100 or IGA score of 0 or 1. [9] The method according to [8] above, wherein the PASI90, PASI100 or IGA score of 0 or 1 is measured 28 weeks after the initial treatment.

[10] The method according to [1] above, wherein the antibody is guselkumab administered subcutaneously.

[11] The method according to any one of the above-mentioned [1] to [3], wherein the antibody is administered subcutaneously at a dose of 25 mg to 200 mg.

[12] The method according to

[11] above, wherein the antibody is administered at a dose of 50 mg or 100 mg.

[13] The method according to any one of the above [1] to [3], wherein the maintenance therapy is administered at an administration interval longer than every 8 weeks.

[14] The method according to

[13] above, wherein the administration interval is selected from the group consisting of 12 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 32 weeks, 36 weeks, 40 weeks, 44 weeks, and 48 weeks.

[15] The method according to any of the above [1] to [3], wherein the maintenance therapy comprises refraining from treatment with an IL-23 antibody.

[16] A method for predicting sustained response to IL-23 antibody therapy and treating a patient with psoriasis, the method comprising: a. administering to said patient an IL-23 antibody comprising a heavy chain variable region amino acid sequence of SEQ ID NO:7 and a light chain variable region amino acid sequence of SEQ ID NO:8 at a dose of 100 mg subcutaneously at an initial dose, 4 weeks after said initial dose, and every 8 weeks after said dose at week 4, wherein said patient is identified as a responder to said IL-23 antibody and has a PASI90, PASI100, or IGA score of 0 or 1; b. obtaining or having obtained a biological sample from said patient; c. At least one indicator from the patient or the biological sample is measured or has been measured at baseline or at any time during treatment, before and during treatment with the IL-23 antibody, selected from the group consisting of clinical parameters including disease duration, BMI and / or IGA score, and PASI, pharmacokinetic markers including blood concentrations of the IL-23 antibody, and serum protein biomarkers including IL17F and MIP1β; d. generating a predictive patient profile based on the measured indicators; and e. administering said maintenance therapy of said IL-23 antibody according to said predicted patient profile, wherein said maintenance therapy is administered at a dosing interval selected from the group consisting of: greater than every 8 weeks, every 12 weeks, every 16 weeks, every 20 weeks, every 24 weeks, every 28 weeks, every 32 weeks, every 36 weeks, every 40 weeks, every 44 weeks, and every 48 weeks.

Claims

[Claim 1] 1. A pharmaceutical composition for use in a method for predicting sustained response to IL-23 antibody therapy and treating a patient with psoriasis, comprising an IL-23 antibody comprising a heavy chain variable region amino acid sequence of SEQ ID NO:7 and a light chain variable region amino acid sequence of SEQ ID NO:8; The method comprises: a. administering the IL-23 antibody subcutaneously to the patient at a dose of 100 mg at an initial dose, 4 weeks after the initial dose, and every 8 weeks after the dose at week 4, wherein the patient is identified as a responder to the IL-23 antibody and has a PASI90, PASI100, or IGA score of 0 or 1; b. obtaining or having obtained a biological sample from said patient; c) measuring or having measured at least one indicator from the patient or the biological sample at baseline or any time during treatment, before and during treatment with the IL-23 antibody, selected from the group consisting of clinical parameters including disease duration, BMI and / or IGA score, and PASI, pharmacokinetic markers including blood concentrations of the IL-23 antibody, and serum protein biomarkers including IL17F and MIP1β; d. generating a predictive patient profile based on the measured indices; e. administering a maintenance therapy of said IL-23 antibody according to said predicted patient profile, wherein said maintenance therapy is administered at a dosing interval selected from the group consisting of: greater than every 8 weeks, every 12 weeks, every 16 weeks, every 20 weeks, every 24 weeks, every 28 weeks, every 32 weeks, every 36 weeks, every 40 weeks, every 44 weeks, and every 48 weeks.

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