TW201307845A - Predictive methods and methods of treating arthritis using IL-17 antagonists - Google Patents

Abstract

The disclosure is directed to novel predictive methods and personalized therapies for treating rheumatoid arthritis (RA). Specifically, the disclosure relates to predicting the likelihood of that a patient having RA will clinically respond to treatment with an an IL-17 binding molecule, e.g., an IL-17 antibody, such secukinumab.

Description

Prediction method and method for treating arthritis by using IL-17 antagonist

The present invention relates to novel predictive methods and personalized therapies for the treatment of rheumatoid arthritis (RA). In particular, the present invention relates to predicting that a patient with RA will have a clinical response to treatment with an IL-17 antagonist (eg, an IL-17 antibody, such as the AIN457 antibody, which is also referred to as "secekumab"). possibility.

The present invention claims the benefit of U.S. Provisional Patent Application Serial No. 61/422,521, filed on Dec.

The human leukocyte antigen (HLA) gene in the major histocompatibility complex (MHC) class II region is the most important genetic risk factor for RA. (Holoshitz (2010) Current Opinion in Rheumatology 22: 293-298). The HLA-DRB1 dual gene encoding a shared epitope (SE) confers a higher risk of RA (Gonzalez-Gay et al., (2002) Sem. Arthritis. Rheum. 31:355-60; Fries et al. (2002) Arthritis and Rheumatism 46:2320-29; van der Helm-van Mil et al., (2006) Arthritis and Rheum. 54:1117-21). SE is an epitope having the amino acid sequence R(Q) ⁷⁰ K/RR AA ⁷⁴ , which is found at positions 70 to 74 in the third hypervariable region of the DRB1 chain. (Feitsma et al., (2010) Arthritis and Rheum. 62:117-25). The HLA-DRB1 dual gene containing SE is prone to cause anti- citrullinated protein antibody (ACPA) + RA disease, and only the SE epitope is associated with ACPA + RA (de Vries et al., (2005) J. Autoimmunity 25: 21 -25). It has been inferred that the citrullylated peptides processed from the guarylated protein (such as citrullinated vimentin) in the synovial fluid of RA patients are recognized by T cells expressing the product of the HLA-DRB1 dual gene containing SE, causing ACPA. Produced (Gregersen et al. (1987) Arthritis Rheum. 30: 1205-13). Thus, SE-containing HLA-DRB1 dual genes are generally thought to contribute to the production of autoantibodies (i.e., ACPA) rather than independently promoting RA progression (van der Helm-van Mil et al., supra). However, recent in vitro studies indicate that SE can act as an immunostimulatory ligand that enhances IL-6 production and Th17 differentiation while inhibiting Treg cell production. (De Almeida et al., (2010) J. Immunol 185: 1927-34). De Almeida et al. showed that SE-activated dendritic cells increased IL-17 production in CD4+ T cells; SE may be associated with direct immune dysregulation via T cell polarization.

Rheumatoid arthritis (RA) is a chronic, inflammatory, systemic autoimmune disease of unknown etiology. It is characterized by symmetric synovitis, which causes cartilage damage and joint destruction and may be complicated by many extra-articular manifestations. If autoantibodies are present, such as rheumatoid factor (RF) and anti-citrullinated protein antibodies (ACPA), RA is considered an autoimmune disease. RA is generally a progressive disease, and functional state decline, significant onset, and premature death can be observed in established RA. The goal of long-term RA treatment is to relieve the disease. Disease-modifying antirheumatic drug (DMARD) is a collection of heterogeneous agents classified according to use and convention, which is the first line treatment for RA patients. The DMARD (most often methotrexate (MTX)) is usually opened before the disease is diagnosed (ie, early RA) before the malformation of the erosive disease and the formed RA. Unfortunately, only about two-thirds of patients respond to DMARDs, and DMARDs only partially control established RA disease. DMARDs also have many adverse effects (such as liver damage, myelosuppression, and severe lung infections), which limits their long-term use. anti-TNF biological agent (Cimzia Enbrel Humira Remicade Simponi For second-line treatment in patients with RA, it is being used in patients with DMARD ineffectiveness and DMARD response. TNF inhibitors are often combined with MTX (or another DMARD) to actively treat established RA. Unfortunately, 30% to 40% of patients with established RA have failed to respond to TNF-α antagonists and most of the patients who initially respond did not achieve complete remission or lost response over time. Has also been concerned with the short-term and long-term tolerability and safety of long-term biotherapeutics, with the greatest attention to serious infections (such as tuberculosis infection) reactivation, hepatotoxicity, increased cardiovascular disease, and induction of myelin loss (or The worsening) and the incidence of malignant diseases increase due to TNF-α antagonism. M. Khraishi (2009) J. Rheumatol Supplement, 82: 25-32; Salliot et al., (2009) Ann. Rheum. Dis. 68: 25-32.

If current RA therapies have the above problems, then there is a need to develop a method of treating RA that first identifies the patient most likely to benefit from the selected RA treatment.

Securkinumab is a novel biological agent for RA in the clinical development stage, which is a high-affinity fully human monoclonal anti-human antibody that inhibits the activity of interleukin-17A. In a RA proof-of-concept (PoC) study, intravenously administered to patients with active RA who received a stable dose of MTX in increasing doses followed by a single dose of 2 (interval 21 days) 1 mg/kg, 3 mg/kg and 10 mg/kg cesikumab. Hueber et al. (2010) Sci. Transl. Med. 2(52): 52-72. The clinical performance of RA in many patients was rapidly improved compared with placebo with serkuumumab. This information demonstrates that neutralizing IL-17A may be effective for RA patients with active RA. However, since patients respond to biological treatments and need to avoid providing drugs to patients who are resistant to them, we have sought a way to treat RA, which first identifies the patients most likely to respond favorably to IL-17. . Thus, the present invention provides novel predictive methods and personalized therapies for the treatment of RA, which maximizes the benefit of antagonizing IL-17 in the RA population by identifying the patients most likely to respond favorably to IL-17 during RA treatment. And to minimize the risk of antagonizing IL-17. The results of this study are based, in part, on the identification of RA patients with dual genes in the dual genome of the SE, HLA-DRB1*SE dual genome or the dual gene in the HLA-DRB1*04 dual genome, ie, IL-17 antagonists (ie, Treatment with securkinumab showed an improved response.

Accordingly, it is an object of the present invention to provide a method for identifying a patient who is likely to respond to treatment of RA with an IL-17 antagonist (e.g., an IL-17 antibody, such as the AINI457 antibody (secekumab)). This is achieved by determining whether the patient has a dual gene in the SE, HLA-DRB1*SE dual genome or a dual gene in the HLA-DRB1*04 dual genome.

Another object of the present invention is to provide a method for determining the likelihood that a RA patient will respond to treatment with an IL-17 antagonist (e.g., an IL-17 antibody, such as cesikumab), by determining whether the patient is This is achieved by having a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome.

Another object of the present invention is to provide a method of treating RA by administering to a patient a therapeutically effective amount of an IL-17 antagonist (e.g., an IL-17 antibody, such as cesikumab), with the proviso that The patient has a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome.

Based on the above objects and findings, various methods are disclosed for predicting the likelihood that a patient with rheumatoid arthritis (RA) will respond to treatment with an IL-17 antagonist (e.g., securizumab). In some embodiments, the methods comprise analyzing the presence or absence of at least one of the dual gene in the SE, HLA-DRB1*04 dual genome, or at least one of the HLA-DRB01*SE dual genome in the biological sample from the patient. Existence (eg, using an automated analyzer) wherein the presence of at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or at least one of the HLA-DRB01*SE dual genome indicates that the patient will be antagonistic to IL-17 Increased likelihood of treatment response, without the presence of at least one dual gene in the SE, HLA-DRB1*04 dual genome, or at least one dual gene in the HLA-DRB01*SE dual genome indicates that the patient will be antagonistic to IL-17 The likelihood of treatment responding is reduced.

Also disclosed herein is a kit for use in the method, for example, for predicting a patient having RA who will be responsive to treatment with an IL-17 antagonist (e.g., sequenumab), for treating a patient a set of patients with RA, and a set comprising at least one capable of detecting the presence of at least one dual gene of the SE, HLA-DRB1*04 dual genome or the HLA-DRB01*SE dual genome or Probes that are not present are used to predict the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist (eg, secukimumab).

Various methods of treating RA are also disclosed herein. In some embodiments, the methods comprise analyzing the presence of at least one dual gene in the SE, HLA-DRB1*04 dual genome, or at least one of the HLA-DRB1*SE dual genome in a biological sample from a RA patient or Not present; and if at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or at least one of the HLA-DRB1*SE dual genome is present in the biological sample, the therapeutically effective amount of the RA patient is administered An IL-17 antagonist, such as cesikumab.

Various methods of selectively treating patients with RA are also disclosed herein. In some embodiments, the methods comprise determining whether a presence or absence of SE in the biological sample from the patient, at least one of the HLA-DRB1*04 dual genome, or at least one of the HLA-DRB01*SE dual genomes Gene; and if at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or at least one of the HLA-DRB01*SE dual genome is present in the biological sample, the therapeutically effective amount of IL is administered to the RA patient -17 antagonist, such as sekkinumab.

Various methods of treating RA patients are also disclosed herein. In some embodiments, the methods comprise receiving at least one of a presence or absence of SE, an HLA-DRB1*04 dual genome, or at least one of the HLA-DRB1*SE dual genomes in the biological sample obtained from the patient. Data for a dual gene; and if the received data indicates that the patient has at least one dual gene in the SE, HLA-DRB1*04 dual genome or at least one dual gene in the HLA-DRB1*SE dual genome, then the A patient with an IL-17 antagonist, such as cesikumab.

Also disclosed herein are various methods of determining the responsiveness of a patient having RA to treatment with an IL-17 antagonist (e.g., sescukib). In some embodiments, the methods comprise analyzing a biological sample from the patient to determine the presence or absence of at least one of the SE, HLA-DRB1*04 dual genome, or at least one of the HLA-DRB1*SE dual genomes. a dual gene; and if at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or the presence of at least one of the dual genes in the HLA-DRB1*SE dual genome is detected, the patient is designated as the pair IL A -17 antagonist (eg, cesikumab) is responsive to treatment.

Also disclosed herein are methods of generating a form of transportable information for predicting the responsiveness of a patient with RA to treatment with an IL-17 antagonist. In some embodiments, the methods comprise analyzing the presence or absence of at least one of the dual genes in the SE, HLA-DRB1*04 dual genome, or at least one of the HLA-DRB1*SE dual genomes in the biological sample from the patient. Exist; and the result of the analysis step is embodied as a form of transportable information.

In some embodiments, the IL-17 antagonist is selected from the group consisting of: a) cesikumab; b) an IL-17 antibody that binds to an IL-17 comprising the following epitope: Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129; c) IL-17 antibodies that bind to the following epitopes in IL-17: Tyr43, Tyr44, Arg46, Ala79, Asp80; d) An IL-17 antibody that binds to an epitope having two IL-17 homodimers of a mature IL-17 protein chain, the epitope comprising Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129 and Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other strand; e) antigen binding to IL-17 homodimer with two mature IL-17 protein chains a base IL-17 antibody comprising Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129 on one strand and Tyr43, Tyr44, Arg46 on the other strand Ala79, Asp80, wherein K IL-17 binding molecule _D of about 100 pM to 200 pM And wherein the in vivo half-life of the IL-17 antagonist is about 4 weeks; and f) an IL-17 antibody comprising an antibody selected from the group consisting of: i) comprising an amino acid as set forth in SEQ ID NO: immunoglobulin heavy chain variable domain sequences (V _H); ii) comprising the SEQ ID NO: immunoglobulin light chain variable domain of 10 amino acid sequence (V _L); iii) comprises SEQ ID NO: 8 shown in the immunoglobulin V _H domain comprising the amino acid sequence and SEQ ID NO: 10 immunoglobulin V _L domain of the amino acid sequences shown; IV) comprising the following high as shown in the immunoglobulin variable regions V _H domain: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; v) comprising an immunoglobulin V _L domain as shown in the following variations of high regions: SEQ ID NO : 4, SEQ ID NO: 5 and SEQ ID NO: 6; vi) an immunoglobulin _VH domain comprising a hypervariable region as shown below: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13; vii) an immunoglobulin _VH domain comprising a hypervariable region as set forth below: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, and comprising a hypervariable region as shown below immunoglobulin V _L domain: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and viii) comprising the Immunoglobulin V _H domain of the hypervariable regions as shown in the following: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, and comprising an immunoglobulin V _L domain as shown in the highly variable region of the SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

In a preferred embodiment, the IL-17 antagonist is an IL-17 binding molecule, preferably a human antibody, most preferably cesikumab.

Other methods, uses, kits, and probes are provided in the following description and in the accompanying claims. Other features, advantages, and aspects of the invention will become apparent to those skilled in the <RTIgt; However, the following description and specific examples are intended to illustrate the preferred embodiments of the invention Various changes and modifications of the present invention will become apparent to those skilled in the <RTIgt;

The term "comprising" encompasses "including" and "consisting of", for example, a composition comprising "including" X may consist solely of X, or may include other items such as X+Y.

The term "administering" with respect to a compound, such as an IL-17 binding molecule or an antirheumatic agent, is used to mean delivery of a compound to a patient by any route.

The phrase "active rheumatoid arthritis" or "active RA" is used to mean RA with visible signs and symptoms (eg, swelling, difficulty in bending, etc.).

The term "analysis" is used to mean the act of identifying, screening, detecting or measuring, which can be performed by any conventional means. For example, the presence of a particular marker in a sample can be analyzed by using an ELISA assay, a northern blot, an imaging method, etc. to detect the presence of a marker in the sample. The terms "analysis" and "assay" encompass the transformation of a substance from one state to another by physical testing of the sample, such as converting a biological sample (eg, a blood sample or other tissue sample) from one state to another. Moreover, the terms "analysis" and "assay" as used herein are used to mean testing and/or measurement. The phrase "analyze from a biological sample of a patient..." and the like is used to mean the presence or absence of a predetermined factor or a specific factor in a sample in a testable (directly or indirectly) sample. It should be understood that where the presence of a substance indicates a probability and the absence of a substance indicates a different probability, the presence or absence of the substance may be used to guide treatment decisions. In the case of HLA typing, assays can be used, such as various well-known typing methods including, for example, serotyping, cell typing, gene sequencing, phenotyping, simple typing, and the like.

An "automatic analyzer" as used herein is any machine that can be used to determine the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome and/or HLA-DRB1*SE dual genome. For example, a PCR machine, an automatic sequencer, a densitometer, a plate reader, a scintillation counter, and the like.

The term "detection" (and similar terms) means the act of extracting specific information from a given source.

The term "about" with respect to the value x means +/- 10% unless the context dictates otherwise. The wording "substantially" does not exclude "completeness". For example, a composition that is "substantially free" of Y may be completely free of Y. The wording "substantially" may be omitted from the definition of the invention as necessary.

"IL-17 antagonist" as used herein refers to the ability to antagonize (eg, reduce, inhibit, reduce, delay) IL-17 function, expression, and/or signaling (eg, by blocking IL-17 binding to IL-) The molecule of the 17 receptor). Non-limiting examples of IL-17 antagonists include IL-17 binding molecules and IL-17 receptor binding molecules. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, an IL-17 antagonist is used.

"IL-17 binding molecule" means any molecule that binds to a human IL-17 antigen either alone or in association with other molecules. Binding reactions can be performed by standard methods (qualitative analysis) (including, for example, binding assays, competitive assays or assays for the inhibition of binding of IL-17 to its receptor, or any type of binding assay), with reference to the use of irrelevant specificity Sexually, but negative control tests for antibodies of the same isotype (eg, anti-CD25 antibodies). Non-limiting examples of IL-17 binding molecules include small molecules such as B cells or fusion tumors, IL-17 receptor decoys and antibodies, and chimeric antibodies, CDR-grafted antibodies or human antibodies, or any fragment thereof, For example, F(ab') ₂ and Fab fragments, as well as single-stranded or single-domain antibodies. Preferably, the IL-17 binding molecule antagonizes (e.g., reduces, inhibits, reduces, delays) IL-17 function, performance, and/or signaling. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, IL-17 binding molecules are used.

"IL-17 receptor binding molecule" means any molecule that binds to the human IL-17 receptor, either alone or in association with other molecules. Binding reactions can be performed by standard methods (qualitative analysis) including, for example, binding assays, competition assays or biological assays for inhibition of binding of IL-17 receptor to IL-17, or any type of binding assay) Negative control tests with antibodies that are irrelevant but of the same isotype (eg, anti-CD25 antibodies) were confirmed. Non-limiting examples of IL-17 receptor binding molecules include small molecules such as B cells or fusion tumors, IL-17 decoys and antibodies to the IL-17 receptor, as well as chimeric antibodies, CDR-grafted antibodies or humans. An antibody, or any fragment thereof, such as F(ab') ₂ and a Fab fragment, and a single chain or single domain antibody. Preferably, the IL-17 receptor binding molecule antagonizes (e.g., reduces, inhibits, reduces, delays) IL-17 function, performance, and/or signaling. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, an IL-17 receptor binding molecule is used.

The term "antibody" as referred to herein includes whole antibodies and any antigen binding portion or single strand thereof. A naturally occurring "antibody" is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as _VH ) and a heavy chain constant region. The heavy chain constant region contains three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as V _L) and a light chain constant region. The light chain constant region contains a domain, CL. V _H and V _L regions can be further subdivided into regions interspersed with the more conserved region (called the framework regions (FR)) the hypervariable region (termed complementarity determining regions (CDR)). Each V _H and V _L is composed of the following sequence from amino-terminus to carboxy-terminus in three CDR's and four FR: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The heavy chain variable region and the light chain variable region contain a binding domain that interacts with the antigen. The constant region of the antibody mediates binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (C1q) of a typical complement system. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, antibodies to the IL-17 or IL-17 receptor are used.

The term "antigen-binding portion" of an antibody as used herein refers to an antibody fragment that retains the ability to specifically bind to an antigen (eg, IL-17). It has been shown that a fragment of a full length antibody can exert the antigen binding function of the antibody. Examples of the binding fragments encompassed within the term "antigen-binding portion" of an antibody include: a Fab fragment which is a monovalent fragment consisting of V _L , V _H , CL and CH1 domains; a F(ab) ₂ fragment which is comprised of bivalent fragment Fab fragments two disulfide bridges in the hinge region of the connection; Fd fragment consisting of the V _H and the CH1 domains; an Fv fragment consisting of the V _L and V _H domains of a single arm of an antibody consisting of; a dAb fragment (Ward et Human, 1989 Nature 341:544-546), which consists of a _VH domain; and an isolated complementarity determining region (CDR). Exemplary antigen binding sites include CDRs ( Table 4 ) as shown in SEQ ID NOS: 1-6 and 11-13 in cesomumab, preferably heavy chain CDR3. Furthermore, although the two domains of the Fv fragment (i.e., V _L and V _H) not encoded by each gene based, but using recombinant methods, by making synthetic linker can be a single protein chain in which the connecting, wherein the V _L Pairing with the _VH region to form a monovalent molecule (referred to as a single chain Fv (scFv); see, eg, Bird et al, 1988 Science 242: 423-426; and Huston et al, 1988 Proc. Natl. Acad. Sci. 85: 5879- 5883). The term "antibody" is also intended to encompass such single chain antibodies. Single-chain antibodies and antigen-binding portions are obtained using conventional techniques known to those skilled in the art. In some embodiments of the disclosed methods, protocols, kits, protocols, uses, and compositions, a single strand of an antibody against IL-17 (eg, cesikumab) or an antibody against the IL-17 receptor is used. Antibody or antigen binding moiety.

The term "pharmaceutically acceptable" means a non-toxic substance that does not interfere with the effectiveness of the biological activity of the active ingredient.

As used herein, "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (eg, an antibody that specifically binds to IL-17 is substantially free of specific binding except for IL-17. Antibody to the antigen). The isolated antibody may be substantially free of other cellular material and/or chemicals. However, an antibody that specifically binds to IL-17 can be cross-reactive with other antigens, such as IL-17 molecules from other species. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, the IL-17 antagonist is an isolated antibody.

The term "monoclonal antibody" or "monoclonal antibody composition" as used herein refers to a preparation of an antibody molecule having a single molecular composition. The monoclonal antibody composition exhibits a single binding specificity and affinity for a particular epitope. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, the IL-17 antagonist is a monoclonal antibody.

The term "human antibody" as used herein is intended to include antibodies in which the framework regions and CDR regions of the variable region are derived from sequences of human origin. "Human antibodies" need not be produced by humans, human tissues or human cells. Human antibodies of the invention may include amino acid residues that are not encoded by human sequences (e.g., mutations introduced in vitro by random or site directed mutagenesis or introduced by somatic mutation in vivo). However, the term "human antibody" as used herein is not intended to include antibodies that have been grafted onto human framework sequences by CDR sequences derived from the germline of another mammalian species, such as a mouse. In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, the IL-17 antagonist is a human antibody.

The term "IL-17" refers to IL-17A (formerly known as CTLA8) and includes polymorphic variants of wild-type IL-17A, IL-17A from various species (eg, humans, mice, and monkeys), and IL. Functional equivalent of -17A. The functional equivalent of IL-17A of the invention preferably has at least about 65%, 75%, 85%, 95%, 96%, 97%, 98% or at least with wild-type IL-17A (eg, human IL-17A). Even 99% overall sequence identity, and substantially retains the ability to induce IL-6 production by human dermal fibroblasts.

The term "K _D" is intended to refer to a specific antibody - antigen interaction of the dissociation rate. As used herein the term "K _D" is intended to refer to the dissociation constant, which is obtained from the Department of K _d to K _a of the ratio (i.e., K _d / K _a), and is expressed in molar concentration (M). K _D values for antibodies can be determined using the generally accepted methods in the art. The method of measuring K _D is to use antibodies surface plasmon resonance biosensor or systems, such as Biacore system. In some embodiments of the invention, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule (eg, IL-17 antibody or antigen binding fragment thereof) binds human IL-17 K _D of about 100 pM to 250 pM.

The term "affinity" refers to the strength of interaction between an antibody and an antigen at a single antigenic site. Within each antigenic site, the variable region of the antibody "arm" interacts with the antigen at multiple sites via weak non-covalent forces; the greater the interaction, the stronger the affinity. Standard assays for assessing the binding affinity of antibodies to IL-17 of various species are known in the art and include, for example, ELISA, Western blotting, and RIA. The binding kinetics of the antibody (e.g., binding affinity) can also be assessed by standard assays known in the art, such as by Biacore analysis. Analysis of the effect of an antibody on the functional properties of IL-17 (e.g., receptor binding, prevention or amelioration of osteolysis) is described in more detail in the Examples.

The terms "individual" and "patient" as used herein include any human or non-human animal. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, and the like.

The antibody "inhibits" one or more of these IL-17 functional properties (eg, biochemical, immunochemical, cellular, physiological, or other biological activity or as determined by methods known in the art and described herein). A similar activity thereof is considered to mean a statistically significant decrease in the specific activity observed for a particular activity relative to the absence of the antibody (or in the presence of a control antibody having an irrelevant specificity). An antibody that inhibits IL-17 activity results in a statistically significant decrease in the measured parameter, such as a decrease of at least 10%, a decrease of at least 50%, 80%, or 90%, and in certain embodiments, the antibody of the invention can inhibit more than 95 %, 98% or 99% of IL-17 functional activity.

Unless otherwise indicated, the term "derivative" is used to define, for example, an IL-17 antagonist of the invention having a specified sequence, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, Sekkingdan Amino acid sequence variants and covalent modifications of an anti-) or IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof). A "functional derivative" includes a molecule having the same qualitative biological activity as the disclosed IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen thereof). A binding fragment, such as cesumuzumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof). Functional derivatives include fragments of IL-17 antagonists and peptide analogs as disclosed herein. A fragment comprises, for example, a region within the sequence of a polypeptide of the invention having the specified sequence. Disclosed herein, the IL-17 antagonist of the functional derivative comprises V _H of the preferred IL-17 binding molecule and / or V _L sequence (e.g. of Table V _H 2 and / or V _L sequences) disclosed herein the disclosed having at least about 65%, 75%, 85%, 95%, 96%, 97%, 98% or even 99% overall sequence identity to the V _H and / or V _L domain, or IL- herein comprising a CDR of a 17 antagonist (eg, cesikumab) having at least about 65%, 75%, 85%, 95%, 96%, 97%, 98%, or even 99% overall sequence identity (eg, with a table) 2. the CDR's 1, 2 or 3 different amino acids), and substantially retains the ability to bind the human IL-6 of IL-17 or IL-17-induced inhibition of e.g. human dermal fibroblast cells.

As used herein, "inhibiting IL-16" refers to the ability of an IL-17 antagonist (eg, cesikumab) to reduce IL-6 production by primary human dermal fibroblasts. Primary human (dermal) fibroblasts produce IL-6 dependent on IL-17 (Hwang et al. (2004) Arthritis Res Ther; 6: R120-128). Briefly, human dermal fibroblasts were stimulated with recombinant IL-17 in the presence of various concentrations of an IL-17 binding molecule having an Fc portion or a human IL-17 receptor. Chimeric anti-CD25 antibody should be used (Basilizimab) was used as a negative control. Supernatants were taken 16 hours after stimulation and analyzed by IL-6 for ELISA. An IL-17 antagonist, such as IL-, as disclosed herein, is tested as described above (i.e., for inhibition of IL-6 production induced by hu-IL-17 in human dermal fibroblasts). 17 binding molecules (eg, IL-17 antibodies or antigen-binding fragments thereof, such as cesikumab) or IL-17 receptor binding molecules (eg, IL-17 antibodies or antigen-binding fragments thereof) inhibit IL-6 production (at 1) nM in the presence of human IL-17) of the IC ₅₀ is typically about 50 nM or less, or 50 nM (e.g. from about 0.01 nM to about 50 nM). In some embodiments of the disclosed methods, protocols, kits, procedures, uses, and compositions, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, Secco secukinumab) or IL-17 receptor binding molecule (e.g. IL-17 as defined antibody or antigen binding fragment thereof) as described above and its functional derivative of the IC IL-6 produced inhibition of ₅₀ 20 nM or about 20 nM More preferably, it is about 10 nM or less, more preferably about 5 nM or less, more preferably about 2 nM or less, more preferably about 1 nM or less.

The term "covalent modification" includes, for example, a polypeptide of the invention having a specified sequence or a fragment thereof, modified with an organic protein or non-protein derivatizing agent, fused to a heterologous polypeptide sequence, and post-translationally modified. Covalently modified, for example, a polypeptide having the specified sequence is still capable of binding to human IL-17 or by inducing IL-17 to induce IL-17 production by human dermal fibroblasts. Covalent modification is conventionally achieved by reacting the targeted amino acid residue with an organic derivatizing agent capable of reacting with a selected side chain or terminal residue, or by utilizing in a recombinant host cell of choice Introduced by the mechanism of post-translational modification. Certain post-translational modifications are the result of the effect of the recombinant host cell on the polypeptide being expressed. The branamine sulfhydryl group and the aspartame sulfhydryl residue are often translated to form the corresponding glutamine sulfhydryl group and the aspartame sulfhydryl residue. Alternatively, the residues are deaminated under slightly acidic conditions. Other post-translational modifications include hydroxylated proline and lysine; phosphorylated serine sulfhydryl, tyrosine or sulfhydryl hydrazide residues; methylated lysine, arginine and histidine Alpha-amino groups of chains, see for example TE Creighton, Proteins: Structure and Molecular Properties, WH Freeman & Co., San Francisco, pp. 79-86 (1983). Covalent modifications include, for example, fusion proteins comprising, for example, a polypeptide of the invention having the specified sequence, and amino acid sequence variants thereof (such as immunoadhesins), and N-terminal fusions to heterologous signal sequences.

The phrase "substantially identical" means that the relevant amino acid or nucleotide sequence (e.g., CDR, V _H or V _L domain) is compared to the reference sequence specific uniform or have insubstantial differences (e.g., except that Conservative amino acid substitution). Non-substantial differences include fewer amino acid changes, such as substitutions at the 1 or 2 of the sequence of 5 amino acids in the designated region. In the case of an antibody, the second antibody has the same specificity and at least 50% of its affinity. Sequences that are substantially identical (e.g., have at least about 85% sequence identity) to the sequences disclosed herein are also part of this application. In some embodiments, the sequence identity can be about 90% or more, such as 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or More than 99%.

"Consistency" with respect to a native polypeptide and its functional derivatives is defined herein as the alignment of the sequences and, if necessary, the introduction of voids to achieve a maximum percent identity, and does not treat any conservative substitutions as sequence identity. The percentage of amino acid residues in a portion of the candidate sequence that are identical to the residues of the corresponding native polypeptide. N- or C-end extensions and insertions should not be considered to reduce consistency. Methods and computer programs for comparison are well known. The percent identity can be determined by a standard alignment algorithm, such as the Basic Local Alignment Search Tool described by Altshul et al. ((1990) J. Mol. Biol., 215: 403 410). , BLAST); Needleman et al, ((1970) J. Mol. Biol., 48: 444 453) algorithm; or Meyers et al, ((1988) Comput. Appl. Biosci., 4: 11 17) algorithm . A set of parameters can be a Blosum 62 scoring matrix with a gap penalty of 12 points, a gap extension penalty of 4 points, and a frame shift gap penalty of 5 points. You can also use the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4:11-17) (which has been incorporated into the ALIGN program (version 2.0)), using the PAM120 weight residue table, 12-point gap length Penalties and a gap penalty of 4 points were used to determine the percent identity between the two amino acid or nucleotide sequences.

"Amino acid" means, for example, all naturally occurring L-alpha-amino acids, and includes D-amino acids. Amino acids are identified by well-known single or three letter names.

The term "amino acid sequence variant" refers to a molecule that differs somewhat in the amino acid sequence compared to the sequences of the invention. For example, amino acid sequence variants of a polypeptide of the invention having a specified sequence are still capable of binding to human IL-17 or, for example, inhibiting IL-17-induced IL-6 production by human dermal fibroblasts. The amino acid sequence variant includes a substitution variant (a variant obtained by removing at least one amino acid residue in the polypeptide of the present invention and appropriately inserting a different amino acid at the same position), an insertion variant a variant (obtained directly adjacent to an amino acid at a specific position by insertion of one or more amino acids in a polypeptide of the invention) and a deletion variant (by removal of the polypeptide of the invention) A variant obtained from one or more amino acids).

"Therapeutically effective amount" as used herein refers to an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule. (eg, an IL-17 antibody or antigen-binding fragment thereof) can be effective to treat, prevent, or prevent a disease, a disease, a cure, a delayed condition, or a relapsed condition after administration to a patient, such as a human patient, in single or multiple doses, Decreasing its severity, improving at least one of its symptoms, or prolonging the survival of the patient over the expected survival period in the absence of the treatment. When applied to an individual active ingredient administered alone (eg, an IL-17 antagonist, such as cesikumab), the term refers to that ingredient alone. When applied to a combination, the term refers to a combined amount of the active ingredient that is administered in combination, continuously or simultaneously, to produce a therapeutic effect.

The term "treatment" or "treat" refers to prophylactic or preventative treatment as well as curative or disease-modifying treatment, including treatment of a patient at risk of contracting a disease or suspected of having an infection. A patient who is ill or has been diagnosed with a disease or medical condition and includes inhibition of clinical relapse. Treatment may be administered to a patient having a medical condition or who may eventually develop the condition to prevent, cure, or relapse the condition, delay its onset, reduce its severity, improve one or more symptoms, or prolong the survival of the patient. And exceeds the expected survival in the absence of this treatment.

The phrase "inflammatory inflammatory arthritis" as used herein refers to a variety of joint conditions involving the immune system and inflammation, and includes autoimmune disorders such as rheumatoid arthritis. Non-limiting examples include seronegative negative spondyloarthropathy such as AS, Reiter's syndrome, PsA, enteropathic arthritis, and other joint diseases such as RA, juvenile rheumatoid arthritis, and systemic disease Rheumatoid arthritis, crystalline arthritis (gout, pseudogout, apatite gout), rheumatic polymyalgia, amyloid arthritis, pigmented villonodular synovitis, synovial chondromatosis, Hemophilic arthritis and reactive synovitis. In some embodiments of the disclosed methods, protocols, uses, kits, and pharmaceutical compositions, the patient has inflammatory arthritis.

The phrase "responsive to treatment" is used to mean that a patient exhibits a clinically meaningful benefit from the treatment after delivery of a particular treatment (eg, an IL-17 binding molecule). In the case of RA, this benefit can be measured using a variety of criteria, such as ACR20, DAS28, ACR50, ACR70, DAS28_CRP, and the like. The phrase "reactive to treatment" is intended to be understood in a comparative manner, not as an absolute response. That is, patients who have a dual gene in the dual genome of the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*04 dual genome ("carrier") are predicted to benefit more from treatment with an IL-17 antagonist. A patient who does not have a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*04 dual genome. These carriers are more responsive to treatment with IL-17 antagonists and may be referred to as "pairs" with the "treatment" of IL-17 antagonists. The term "reactivity" is a measure of the level of therapeutic benefit.

The phrase "receiving data" is used to mean ownership of information by any available means, such as verbal, electronic (eg, by e-mail, encoded on a magnetic disk or other medium), written, and the like.

"Rheumatoid arthritis" or "RA" refers to chronic systemic inflammatory arthritis that may affect multiple tissues and organs but primarily invades the synovial joints. The 2010 ACR/EULAR guidelines (see Aletaha et al., (2010) Ann. Rheum. Dis. 69: 1580-1588) can be used to classify patients as having RA.

"C-reactive protein" and "CRP" refer to serum C-reactive protein, which is used as an indicator of acute phase response to inflammation. The content of CRP in plasma can be given in any concentration (for example, mg/dl, nmol/L). The amount of CRP can be measured by a variety of well-known methods, such as radiation immunodiffusion, electro-immunoassay, immunoturbidimetric assay, ELISA, turbidity assay, fluorescence polarization immunoassay, and laser nephelometry. The CRP test can use a standard CRP test or a highly sensitive CRP (hs-CRP) test (i.e., a high sensitivity test capable of measuring the low CRP content in a sample using a laser turbidimetric method). Kits for detecting CRP content are commercially available from various companies such as Calbiotech Inc., Cayman Chemical, Roche Diagnostics Corporation, Abazyme, DADE Behring, Abnova Corporation, Aniara Corporation, Bio-Quant Inc., Siemens Healthcare Diagnostics, and the like.

"High CRP content" means a normal CRP content as defined in the 2010 ACR/EULAR guidelines (Aletaha et al. (2010) Ann. Rheum. Dis. 69: 1580-88). According to the 2010 ACR/EULAR guidelines, normal/abnormal CRP is based on local laboratory standards. Each local laboratory will use an abnormal (high) CRP cutoff based on the laboratory's rules for calculating normal maximum CRP. The physician generally instructs the local laboratory to perform a CRP test, and the local laboratory uses the specific laboratory to calculate normal CRP rules reporting normal or abnormal CRP (low or high). Therefore, "high CRP content" as used herein is not intended to mean a particular value unless the context dictates otherwise, as the normal CRP value will vary from laboratory to laboratory. "hsCRP" refers to the amount of CRP in the blood as measured by the highly sensitive CRP test.

"Red blood cell sedimentation rate", "ESR", "sedimentation rate" and "sedrate" refer to the sedimentation rate of red blood cells in a patient sample. ESR reflects the presence of plasma viscosity and acute phase proteins and is usually reported in "mm/hr". ESR is known by measuring the distance that red blood cells precipitate in the tube over time. A typical ESR test uses the Weiss test, the ZSR test, and the Wintrobe test. (Moseley and Bull (1982) Clin. Lab Haematol. 4: 169-78; Miller et al., (1983) Br Med J (Clin Res Ed) 286 (6361): 266; Wetteland P et al., (1996) J. Intern. Med. 240(3): 125-310, which is incorporated herein by reference in its entirety. Commercial kits for measuring ESR are available, for example, from ARKRAY USA, BD Diagnostic Systems, and Poly Med Co. Inc. ESR instruments are found, for example, in U.S. Patent 6,974,701, and are commercially available from various companies, such as Steelex Scientific, Nicesound Electronics Co., Globe Scientific Inc., Alifax, Analysis Instruments AB, Streck Laboratories, PolyMed Co. Inc., and Quantimetrix.

"High ESR" means a normal ESR as defined in the 2010 ACR/EULAR guidelines (Aletaha et al. (2010) Ann. Rheum. Dis. 69: 1580-88). According to the 2010 ACR/EULAR guidelines, normal/abnormal ESR is based on local laboratory standards. Each local laboratory will use an abnormal (high) ESR cutoff based on the laboratory's calculation of the normal maximum ESR. Physicians generally instruct local laboratories to perform ESR tests, and local laboratories use the laboratory to calculate normal ESR regulations to report normal or high ESR. Therefore, "high ESR" as used herein is not intended to mean a particular value unless the context dictates otherwise, as the normal ESR value will vary from laboratory to laboratory.

"Rheumatoid factor" or "RF" refers to an autoantibody to the Fc portion of an IgG antibody, which is often present in RA patients. "RF" as used herein includes any RF isotype, such as IgG, IgE, IgM, and IgA. RF can be analyzed using a variety of well known techniques that can be used to determine the presence or absence of a particular antibody, such as ELISA assays, agglutination assays, turbidimetric assays, and the like. The RF content can be measured by the laboratory in a variety of ways (eg IU/ml, units/ml and potency (using a dilution test to measure the extent to which the patient's blood sample can be diluted before RF can no longer be detected, eg 1:80) The potency is reported as compared to the 1:20 titer indicating that the RF is more detectable). RF kits are commercially available, for example, from IBL-America (Immuno-Biological Laboratories).

A patient with a positive RF seropositivity is referred to herein as "RF+." Similarly, if the sample from the patient contains RF, the sample is "RF+". Each local laboratory will use the normal RF content cutoff based on the laboratory's calculation of normal maximum RF. As suggested by Aletaha et al. (2010) Ann. Rheum. Dis. 69:1580-1588, patients will be considered to be RF+ based on the upper limit of normal values [ULN] for each laboratory test and analysis; The value of the ULN tested and analyzed by the respective laboratory is RF+. Therefore, "RF+" as used herein is not intended to mean a particular value unless the context dictates otherwise, as the ULN will vary from laboratory to laboratory. As a non-limiting example, the normal range of RF in laboratory X for a given blood is 14 to 60 units/ml at the time of testing. At the time of testing, the normal range of RF in the laboratory Y established blood is 40 IU/ml. At the time of testing, the normal range of RF in laboratory Z for established blood ranged from 1:20 to 1:80 titer. Therefore, if Laboratory X reports an RF content greater than 60 units/ml, if Laboratory Y reports an RF value greater than 40 IU/ml, or if Laboratory Z reports an RF titer greater than 1:80, the patient is RF+.

The term "seropositive" is used to mean the presence of a particular substance (eg, RF) in the patient's serum. The term "serotype" is used to mean that a particular serological antigen (eg, HLA-DR4 serological antigen) of a particular patient is seropositive.

"Anti-citrullinated protein antibody", "ACPA", "anti-cyclic citrullinated peptide antibody" and "anti-CCP antibody" refer to an autoantibody that binds to a guanosylated amino acid residue on a protein, which is found in RA. In the joint of the patient. Cyclic citrullylated peptides are used in in vitro assays (eg, ELISA assays) to determine the presence of ACPA in the blood of a patient; thus, ACPA is also referred to as an "anti-CCP" antibody. The ACPA content can be analyzed using a variety of well known techniques that can be used to determine the presence or absence of a particular antibody, such as agglutination, ELISA analysis, and the like. ACPA kits are commercially available, such as DIASTAT from Axis-Shield Diagnostics, Ltd. (UK). Anti-CCP antibody test and AxSYM Anti-CCP from Abbot Diagnonstics (Germany) Set of groups.

Patients with positive ACPA seropositivity are referred to herein as "ACPA+." Similarly, if the sample from the patient contains ACPA, the sample is "ACPA+". Each local laboratory will use the cutoff value for normal ACPA content based on the laboratory's calculation of the normal maximum ACPA. As suggested by Aletaha et al. (2010) Ann. Rheum. Dis. 69:1580-1588, patients will be considered to be ACPA+ based on the upper limit of normal values [ULN] for each laboratory test and analysis; The value of the ULN tested and analyzed by the respective laboratory is ACPA+. Therefore, "ACPA+" as used herein is not intended to mean a particular value unless the context dictates otherwise, as the ULN will vary from laboratory to laboratory. As a non-limiting example, the reference range for ACPA in laboratory A established blood is <20 EU (any ELISA unit) at the time of testing. At the time of testing, the reference range for ACPA in laboratory B established blood was <5 U/ml. Therefore, if laboratory A reports an ACPA value greater than 20 EU or if laboratory B reports an ACPA value greater than 5 U/ml, the patient is ACPA+.

The normal/abnormal and reference ranges for optional ACPA, ESR, RF and CRP can be found, for example, in Fischbach and Dunning (2009) "A Manual of Laboratory and Diagnositc Tests" (8th Edition) in Wolters Kluwer/Lippincott Williams and Williams. This is incorporated herein by reference.

The phrase "high risk RA patient" is used to define the following patients: a) both RF+, ACPA+ or both RF+ and ACPA+; and b) with high CRP content, high ESR or high CRP content, and high ESR. In some embodiments of the disclosed methods, uses, compositions, kits, and the like, the patient is a high risk RA patient.

"Selection" and "selection" as used herein with respect to a patient are meant to specifically select a particular patient from a larger group of patients because the particular patient meets predetermined criteria, such as the patient having SE, HLA-DRB1* 04 dual gene or HLA-DRB1*SE dual gene. Similarly, "selective treatment of a patient with RA" provides treatment to a RA patient that is specifically selected from a larger group of patients, as the particular patient meets predetermined criteria, such as the patient having SE, HLA-DRB1 *04 dual gene or HLA-DRB1*SE dual gene.

"Predict" as used herein indicates that the method as described herein provides information to enable a health care provider to determine that an individual having RA will respond to treatment with an IL-17 binding molecule or will be treated with an IL-17 binding molecule. The possibility of a more favorable reaction. It does not refer to the ability to predict the response with 100% accuracy. In fact, the skilled artisan will understand that it refers to the increased probability.

As used herein, "Possibility" and "Possible" are measures of the probability of an event occurring. It is used interchangeably with "probability." Possibility refers to a probability greater than speculation but less than a certain probability. Therefore, if a reasonable person uses common sense, training, or experience to conclude that a certain situation is likely to occur, the event is likely to occur. In some embodiments, once the likelihood is determined, the patient can be treated with IL-17 binding molecules (or continue treatment, or treated at an increased dose), or the patient may not be treated with IL-17 binding molecules (or discontinued treatment, or Treatment at a reduced dose).

The term "obtaining" means obtaining, for example, obtaining ownership in any way.

The phrase "increased probability" refers to an increase in the probability of an event occurring. For example, the methods herein allow predictive patients to respond to IL-17 binding molecule therapy compared to RA patients who do not have the SE or HLA-DRB1*04 dual genome or the dual gene in the HLA-DRB1*SE dual genome. Whether the likelihood increases or increases the likelihood of a better response to IL-17 binding molecule therapy.

The phrase "further increase in likelihood" means that the likelihood of an increase has increased. The data disclosed herein indicates the additive effect of the HLA-DRB1*04 dual gene. Additional information disclosed herein indicates the additive effect of HLA-DRB1*SE dual genes. Therefore, individuals with one HLA-DRB1*04 dual gene may be more likely to respond to treatment with IL-17 binding molecules, while individuals with two HLA-DRB1*04 dual genes are treated with IL-17 binding molecules. The possibility of a reaction may increase further (additiveness). In addition, individuals with one HLA-DRB1*SE dual gene may be more likely to respond to IL-17 binding molecule therapy, whereas individuals with two HLA-DRB1*SE dual genes are treated with IL-17 binding molecule The possibility of a reaction may increase further (additiveness).

The phrase "reduced likelihood" refers to a decrease in the probability of an event occurring. For example, the methods herein allow for the prediction of IL-17 binding molecules in RA patients compared to the dual gene in the dual genome of the SE or HLA-DRB1*04 dual genome or the dual gene in the HLA-DRB1*SE dual genome. Whether the likelihood of treatment is reduced or whether the likelihood of a better response to IL-17 binding molecule therapy is reduced.

"HLA" refers to human leukocyte antigen. HLA is located on chromosome 6p21.31 and includes a region of approximately 3.6 Mbp, depending on the simplex. HLA molecules are encoded by three sets of genes: a class I HLA gene, a class II HLA gene, and a class III HLA gene. Class I HLA proteins are encoded by the genes HLA-A, HLA-B and HLA-C. Class II HLA protein lines are encoded by the following genes: HLA-DR, HLA-DQ, HLA-DP, HLA-DM, HLA-DOA, and HLA-DOB. Class II HLA proteins belong to the complement system. Polymorphic Class I HLA genes HLA-A, HLA-B and HLA-C and Class II HLA genes HLA-DR, HLA-DQ and HLA-DP encode various proteins (see eg hla.alleles.org/proteins/class2) .html) and various antigens (see eg hla.alleles.org/antigens/recognised_serology.html).

Class II HLA molecules consist of two transmembrane polypeptides (alpha and beta chains). The beta strand is more pleomorphic than the alpha strand and is typically HLA typed for the beta strand (eg, HLA-DRB1 to DRB9). HLA dual genes were named according to the 2010 WHO Nomenclature Committee for Factors of the HLA System (Marsh et al., (2010) Tissue Antigens 75:291-455; Marsh et al., (2010) Bone Marrow Transplantation 45: 846-8). Several digits are used to identify HLA dual genes. Specific HLA loci (HLA-A, HLA-B, HLA-C, HLA-DR, HLA-DQ, and HLA-DP) are separated by a symbol * from two digits that specify the serological equivalent of the antigen. (This level describes "type" or "dual genome"). For example, HLA-DRB1*04 represents the dual genome from the HLA-DRB1 locus. This "two-digit" resolution indicates a dual genome consisting of various dual genes encoding similar antigens (eg, HLA-DR4 serological antigens) or consensus high sequence homology (eg, the dual genome from the HLA-DRB1 locus). This is followed by a colon and two or more digits that identify a particular encoded protein (this level describes "subtypes" or "dual gene subtypes"). For example, HLA-DRB1*04:01 is a specific dual gene encoding an HLA-DRβ chain with a specific amino acid sequence in the HLA-DRB1*04 dual genome. This "quadraran" resolution indicates a particular genomic sequence variation within the dual genome that results in a difference in the amino acid sequence of the encoded polypeptide product. The dual gene may be further defined using additional colons and numbers indicating that the synonymous DNA within the coding region of the dual gene is substituted or indicates a DNA difference in the non-coding region (9 digit level).

"HLA-DRB1*04 dual genome" refers to a dual genome (or type) consisting of various dual genes encoding HLA-DR4 serological antigens or shared high sequence homology from the HLA-DRB1 locus.

"HLA-DRB1*04 dual gene" or "dual gene in HLA-DRB1*04 dual genome" refers to a dual gene in the HLA-DRB1*04 dual genome, such as HLA-DRB1*04:01, HLA-DRB1*04 :05 and so on. An exemplary HLA-DRB1*04 dual gene IMG/HLA database (part of the EMBL-EBI database) reference number is shown in Table 1 , the sequence of which can be accessed via www.ebi.ac.uk/imgt/hla/ Nomenclature/index.htmlGet .

Table 1 : IMG/HLA database reference numbers for HLA-DRB1*04 dual genes. A new HLA-DRB1*04 dual gene may be discovered and this list is not exhaustive.

The phrase "at least one dual gene" is used in the previous description of "a dual gene in the HLA-DRB1*04 dual genome" to refer to the HLA-DRB1*04 dual gene. These sequences are incorporated herein by reference in their entirety.

The "product of the HLA-DRB1*04 dual gene" includes nucleic acid products of the HLA-DRB1*04 dual gene, such as mRNA, microRNA, RNA fragment, and the like, and the polypeptide product of the HLA-DRB1*04 dual gene. The "polypeptide product of the HLA-DRB1*04 dual gene" refers to a polypeptide encoded by the HLA-DRB1*04 dual gene, a polypeptide fragment encoded by the HLA-DRB1*04 dual gene, and an HLA-DR4 serological antigen. "Nucleic acid product of HLA-DRB1*04 dual gene" refers to any RNA product of HLA-DRB1*04 dual gene and fragments thereof.

"HLA-DR4 serotype" refers to the serotype of a patient who exhibits a polypeptide product of the HLA-DRB1*04 dual gene (eg, HLA-DR4 serological antigen).

"Shared epitope" or "SE" is an antigen having amino acid sequence R/Q ⁷⁰ K/RRAA ⁷⁴ (SEQ ID NO: 18) found at positions 70 to 74 in the third hypervariable region of the DRB1 chain. Decide on the basis. It is currently known that consensus epitopes are presented by the following dual genes: HLA-DRB1*01:01, HLA-DRB1*01:02 and HLA-DRB1*01:04 (referred to as HLA-DRB1*01 SE dual genome); HLA -DRB1*04:01 (formerly known as Dw4 type), HLA-DRB1*04:04, HLA-DRB1*04:05, HLA-DRB1*04:08, HLA-DRB1*04:09, HLA-DRB1* 04:10, HLA-DRB1*04:13, HLA-DRB1*04:16, HLA-DRB1*04:19, HLA-DRB1*04:21 (referred to as HLA-DRB1*04 SE dual genome); HLA- DRB1*10:01; HLA-DRB1*13:03; HLA-DRB1*14:02 and HLA-DRB1*14:06 (referred to as HLA-DRB1*14 SE dual genome). Gorman and Criswell (2002) Genetics 28: 59-77. A classification system for the risk of RA conferred by HLA-DRB1 dual genes, including the dual gene encoding the polypeptide product with SE, can be found in J. Imboden (2009) Annu. Rev. Pathol. Mech. Dis. 4:417-34 This document is incorporated herein by reference in its entirety. Additional information on SE can be found in Holokitz (2010) Current Opinion in Rheumatology 22: 293-298.

As used herein, "HLA-DRB1*SE dual genome" refers to a dual genome (or type) consisting of the following dual genes from the HLA-DRB1 locus: HLA-DRB1*01:01, HLA-DRB1*01:02, HLA-DRB1*04:01, HLA-DRB1*04:04, HLA-DRB1*04:05, HLA-DRB1*04:08, HLA-DRB1*10:01 and HLA-DRB1*14:02. It is worth noting that some of the dual genes in the HLA-DRB1*SE dual genome are also in the HLA-DRB1*04 dual genome. Thus, in some cases, detection of a dual gene in the HLA-DRB1*SE dual genome can also allow detection of a dual gene in the HLA-DRB1*04 dual genome (and vice versa). An IMG/HLA database reference number for an exemplary HLA-DRB1*SE dual gene is available at www.ebi.ac.uk/imgt/hla/nomenclature/index.html . "HLA-DRB1*SE dual gene" or "dual gene in HLA-DRB1*SE dual genome" refers to a dual gene in the HLA-DRB1*SE dual genome, such as HLA-DRB1*01:01, HLA-DRB1*01 :02. The phrase "at least one dual gene" is used to refer to the "additional gene in the HLA-DRB1*SE dual genome" to refer to the HLA-DRB1*SE dual gene.

The "product of the HLA-DRB1*SE dual gene" includes nucleic acid products of the HLA-DRB1*SE dual gene, such as mRNA, microRNA, RNA fragment, and the like, and the polypeptide product of the HLA-DRB1*SE dual gene. The "polypeptide product of the HLA-DRB1*SE dual gene" refers to a polypeptide encoded by the HLA-DRB1*SE dual gene, a polypeptide fragment encoded by the HLA-DRB1*SE dual gene, and SE. "Nucleic acid product of HLA-DRB1*SE dual gene" refers to any RNA product of HLA-DRB1*SE dual gene and fragments thereof.

"HLA-DRSE serotype" refers to the serotype of a patient who exhibits a polypeptide product of the HLA-DRB1*SE dual gene.

An "equivalent genetic marker" of a related dual gene (eg, an HLA-DRB1*04 dual gene or an HLA-DRB1*SE dual gene) refers to a genetic marker associated with a related dual gene, eg, which exhibits a linkage disequilibrium or is associated with a dual The genetic linkage of genes. An equivalent genetic marker can be used to determine if a patient has a dual gene in the SE, HLA-DRB1*04 dual genome and/or a dual gene in the HLA-DRB1*SE dual genome.

The term "probe" refers to any substance that is suitable for the specific detection of another substance (eg, a substance related to a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome). . The probe may be an oligonucleotide or a binding oligonucleotide that specifically hybridizes to a specific region within the HLA-DRB1*04 dual gene or HLA-DRB1*SE dual gene. A conjugated oligonucleotide refers to an oligonucleotide (e.g., an antibody specific for an antigen) covalently bound to a chromophore or a ligand-containing molecule (e.g., an antigen) and highly specific to the receptor molecule. The probe may also be a PCR primer that, along with another primer, is used to amplify a particular region within the HLA-DRB1*04 dual gene or HLA-DRB1*SE dual gene. Furthermore, the probe may be an antibody that specifically recognizes the HLA-DRB1*04 dual gene or the HLA-DRB1*SE dual gene or the HLA-DRB1*04 dual gene or the HLA-DRB1*SE dual gene polypeptide product or serological antigen. Furthermore, the probe may be any substance capable of detecting the equivalent genetic marker of the HLA-DRB1*04 dual gene or the HLA-DRB1*SE dual gene.

As used herein, "genomic sequence" refers to a DNA sequence that is present in the genome and includes a region within the dual gene, the dual gene itself, or a larger DNA sequence containing the chromosome of the related dual gene.

The term "biological sample" as used herein refers to a sample from a patient that can be used for identification, diagnosis, prediction or monitoring purposes. Preferred test samples include synovial fluid, blood, blood derived products (such as leukocytes, serum and plasma), lymph, urine, tears, saliva, cerebrospinal fluid, buccal swab, sputum or tissue samples. In addition, those skilled in the art will appreciate that certain test samples should be easier to analyze after partial isolation or purification procedures (eg, isolation of DNA from whole blood).

"Oligonucleotide" refers to a short nucleotide sequence, for example 2 to 100 bases.

The term "capable" is used to mean the ability to achieve a given result, for example, the ability of a probe to detect the presence of a particular substance means that the probe is capable of detecting that particular substance.

The phrase "previously treated for RA" and "previous RA treatment" and the like are used to mean that the patient has previously been treated with an anti-rheumatic agent for RA therapy, for example, a patient with prior RA therapy, anti-rheumatic agent or treatment. Invalid programs, underreacted, or intolerant to previous RA therapies, antirheumatic agents, or treatment regimens. Such patients include patients previously treated with MTX, DMARD and/or biological agents (such as TNFα antagonists). The phrase "previously not treated for RA" is used to mean that the patient has not previously been treated with an anti-rheumatic agent for RA, ie, the patient is "untreated."

The term “invalid” in previous RA therapy means: (1) the patient has no meaningful clinical benefit (initially lacking efficacy); (2) the patient has a measurable and meaningful response, but for it, the response should be better, eg Achieve low RA disease activity or RA remission (also known as "underreaction"); (3) patients with deterioration after initial excellent response (secondary loss of efficacy); and (4) patients with excellent response, but due to side effects And stop (also known as "intolerance"). Patients demonstrating insufficient TNF response (TNF-IR) or intolerance to TNF should be considered ineffective for TNF. Patients exhibiting insufficient methotrexate response (MTX-IR) or intolerance to MTX should be considered ineffective for MTX. Patients presenting DMARD-insensitive (DMARD-IR) or intolerant to DMARD should be considered ineffective as DMARD.

The phrase "treatment regimen" means a mode of treating a disease, such as the mode of administration used during the treatment of RA. The treatment regimen can include an induction regimen and a maintenance regimen.

The phrase "induction regimen" or "induction period" refers to a treatment regimen (or part of a treatment regimen) for initial treatment of a disease. In some embodiments, the disclosed methods, uses, kits, procedures, and protocols (eg, methods of treating inflammatory arthritis (eg, RA, such as high-risk RA patients) use an induction regimen. The general goal of the induction regimen is to provide a high dose of the drug to the patient during the initial period of the treatment regimen. The induction regimen may (partly or once) administer a dose greater than the dose used by the physician during the maintenance regimen to administer the drug at a frequency that is higher than the frequency at which the physician administers the regimen during the maintenance regimen, or both. In some embodiments of the disclosed methods, uses, kits, procedures, and protocols, the inducing dose can be delivered in a single high dose infusion (e.g., about 30 mg/kg) during the induction regimen. Alternatively, the inducing dose (e.g., about 10 mg/kg) can be delivered in a number of (e.g., two or three) infusions. Alternatively, the inducing dose (e.g., about 75 to 300 mg) can be delivered in a number of subcutaneous injections. The drug can be delivered via the subcutaneous (sc) route during the induction regimen, for example, subcutaneously delivering a dose of from about 75 mg to about 300 mg (eg, subcutaneous delivery of about 75 mg, subcutaneous delivery of about 150 mg, subcutaneous delivery of about 300 mg), or via vein Internal (iv) route delivery of the drug, for example, intravenous delivery of a dose of from about 1 mg/kg to about 30 mg/kg (eg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, about 30 mg/kg) Or deliver the drug via any other route of administration (eg intramuscular im). In some embodiments of the disclosed methods, compositions, kits, uses, and protocols, an IL-17 antagonist (eg, cesikumab) is delivered by intravenous administration during at least a portion of the induction regimen. In some embodiments, the induction regimen comprises administering an IL-17 antagonist at a dose of from about 1 mg/kg to about 30 mg/kg, from about 1 mg/kg to about 10 mg/kg, preferably about 10 mg/kg. (eg Sekerkinumab). In other embodiments, the inducing dose is delivered once a week, once every two weeks, every other week, or once a month, preferably every other week. In other embodiments, the induction regimen uses from 1 to 10 doses of an IL-17 antagonist (eg, cesikumab), preferably three doses of an IL-17 antagonist (eg, cesikumab).

The phrase "maintenance regimen" or "maintenance period" refers to a treatment regimen (or part of a treatment regimen) that is used to maintain a patient during treatment of a disease, such as maintaining a patient's remission for a longer period of time (months or years). In some embodiments, the disclosed methods, uses, and protocols (eg, methods of treating inflammatory arthritis (eg, RA, such as high-risk RA patients)) use a maintenance regimen. The maintenance regimen may use continuous therapy (eg, administration of the drug at regular intervals, such as once a week, once a month, once a year, etc.), or intermittent therapy (eg, discontinued treatment, intermittent treatment, treatment at relapse, or Treatment after reaching certain predetermined criteria [eg pain, disease performance, etc.]. The drug can be delivered via the subcutaneous route during the maintenance regimen, for example, subcutaneously delivering a dose of from about 75 mg to about 300 mg (eg, subcutaneous delivery of about 75 mg, subcutaneous delivery of about 150 mg, subcutaneous delivery of about 300 mg), or delivery via an intravenous route. The drug, for example, intravenously delivers a dose of from about 1 mg/kg to about 30 mg/kg (eg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, about 30 mg/kg) or via any other administration. Routes (eg, intramuscular im) deliver drugs. In some embodiments of the disclosed methods, uses, and protocols, an IL-17 antagonist (eg, cesikumab) is delivered by subcutaneous administration during a maintenance regimen. In some embodiments, the maintenance regimen comprises administering an IL-17 antagonist at a dose of from about 75 mg to about 300 mg, from about 75 mg to about 150 mg, preferably about 75 mg or about 150 mg (eg, Sekukindan anti). In some embodiments, the maintenance regimen comprises administering a dose of an IL-17 antagonist (eg, cesikumab) on a monthly basis.

The phrase "a member for administration" is used to indicate any available device for administering a drug to a patient, including, but not limited to, a prefilled syringe, vial and syringe, injection pen, autoinjector, intravenous drip. Injectors and bags, pumps, etc. Using these items, the patient can self-administer the drug (ie, to administer the drug for himself) or the physician can administer the drug.

The phrase "specific hybridization" is used to mean specific binding under stringent hybridization conditions. Stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Aqueous and non-aqueous methods are described in this reference and either can be used. An example of stringent hybridization conditions is hybridization in 6X sodium chloride/sodium citrate (SSC) at about 45 °C followed by at least one wash in 0.2X SSC, 0.1% SDS at 50 °C. A second example of stringent hybridization conditions is hybridization in 6X SSC at about 45 °C followed by at least one wash in 0.2 x SSC, 0.1% SDS at 55 °C. Another example of stringent hybridization conditions is hybridization in 6X SSC at about 45 °C followed by at least one wash in 0.2X SSC, 0.1% SDS at 60 °C. Another example of stringent hybridization conditions is hybridization in 6X SSC at about 45 °C followed by at least one wash in 0.2X SSC, 0.1% SDS at 65 °C. Highly stringent conditions include hybridization in 0.5 M sodium phosphate, 7% SDS at 65 °C, followed by washing at 65 °C in 0.2 x SSC, 1% SDS at least once.

The phrase "region of nucleic acid" is used to indicate a smaller sequence within a larger nucleic acid sequence. For example, a gene is a region of a chromosome, an exon is a region of a gene, and the like.

Various aspects of the invention are described in more detail in the following subsections. All patents, patent applications, references, and other publications are hereby incorporated by reference.

IL-17 antagonist and pharmaceutical composition

Various disclosed pharmaceutical compositions, protocols, procedures, uses, methods, and kits use IL-17 antagonists, such as IL-17 binding molecules (eg, IL-17 antibodies or antigen-binding fragments thereof, such as cesikumab) Or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof).

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), comprises at least one CDR1, CDR2, and CDR3 comprising a hypervariable region in turn. An immunoglobulin heavy chain variable domain ( _VH ) having the amino acid sequence SEQ ID NO: 1 (NYWMN) having the amino acid sequence SEQ ID NO: 2 (AINQDGSEKYYVGSVKG), and the CDR3 has Amino acid sequence SEQ ID NO: 3 (DYYDILTDYYIHYWYFDL).

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), comprises at least one CDR1', CDR2' comprising a hypervariable region in turn. and CDR3 'of an immunoglobulin light chain variable domain (V _L), the CDR1' having the amino acid sequence of SEQ ID NO: 4 (RASQSVSSSYLA) , the CDR2 'having the amino acid sequence of SEQ ID NO: 5 (GASSRAT) And the CDR3' has the amino acid sequence SEQ ID NO: 6 (QQYGSSPCT).

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), comprises at least one CDR1-x, CDR2 comprising a hypervariable region in turn. An immunoglobulin heavy chain variable domain ( _VH ) of -x and CDR3-x having the amino acid sequence SEQ ID NO: 11 (GFTFSNYWMN) having the amino acid sequence SEQ ID NO : 12 (AINQDGSEKYY), and the CDR3-x has the amino acid sequence SEQ ID NO: 13 (CVRDYYDILTDYYIHYWYFDL-WG).

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab), comprises at least one immunoglobulin _VH domain and at least one immunization a globulin V _L domain, wherein: a) the immunoglobulin V _H domain comprises: i) a hypervariable region CDR1, CDR2 and CDR3, the CDR1 having the amino acid sequence SEQ ID NO: 1, the CDR2 having an amino acid sequence SEQ ID NO: 2, and the CDR3 has the amino acid sequence SEQ ID NO: 3; or ii) the hypervariable region CDR1-x, CDR2-x and CDR3-x, the CDR1-x having the amino acid sequence SEQ ID NO : 11, CDR2-x having the amino acid sequence of SEQ ID NO: 12, and CDR3-x having the amino acid sequence of SEQ ID NO: 13; and b) the immunoglobulin V _L domain comprises hypervariable regions CDR1 'CDR2' and CDR3', the CDR1' having the amino acid sequence SEQ ID NO: 4, the CDR2' having the amino acid sequence SEQ ID NO: 5, and the CDR3' having the amino acid sequence SEQ ID NO: 6 or Its direct CDR' equivalent.

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab), comprises at least one immunoglobulin _VH domain and at least one immunization a globulin V _L domain, wherein: a) the at least one immunoglobulin _VH domain comprises, in order: i) a hypervariable region CDR1, CDR2 and CDR3, the CDR1 having an amino acid sequence of SEQ ID NO: 1, the CDR2 having an amine a base acid sequence of SEQ ID NO: 2, and the CDR3 has the amino acid sequence SEQ ID NO: 3; or ii) the hypervariable region CDR1-x, CDR2-x and CDR3-x, the CDR1-x having an amino acid sequence SEQ ID NO: 11, CDR2-x having the amino acid sequence of SEQ ID NO: 12, and CDR3-x having the amino acid sequence of SEQ ID NO: 13; and b) at least one immunoglobulin V _L domain sequence Included in the hypervariable region CDR1', CDR2' and CDR3', the CDR1' having the amino acid sequence SEQ ID NO: 4, the CDR2' having the amino acid sequence SEQ ID NO: 5, and the CDR3' having an amino acid sequence SEQ ID NO: 6 or its direct CDR' equivalent.

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) comprises: a) comprises as set forth in SEQ ID NO:8 immunoglobulin heavy chain variable domain amino acid sequence (V _H); b) comprising the SEQ ID NO: immunoglobulin light chain variable domain amino acid sequence (V _L) shown in FIG. 10; c ) comprising the SEQ ID NO: FIG. 8 of the immunoglobulin V _H domain comprising the amino acid sequence and SEQ ID NO: immunoglobulin V _L domain of the amino acid sequence shown in FIG. 10; d) comprising the less immunoglobulin V _H domain shown in high hypervariable regions: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; e) comprising an immunoglobulin V _L domain as shown in the highly variable region of the SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; f) an immunoglobulin _VH domain comprising a hypervariable region as shown below: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13; g) an immunoglobulin _VH domain comprising a hypervariable region as set forth below: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and comprising as shown below V _L domain of immunoglobulin hypervariable regions: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; or h) package Immunoglobulin V _H domain as shown in the hypervariable regions of the following: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, and comprising an immunoglobulin V _L as shown in the highly variable region of the Domain: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

For ease of reference, the amino acid sequence based on the Kabat definition and the hypervariable region of the cetuzumab monoclonal antibody as determined by X-ray analysis and using the method of Chothia and co-workers is provided in Table 2 below.

Table 2 : Amino acid sequence of the hypervariable region of the single antibody of cesomumab. The amino acid highlighted in bold is part of the CDR loop, while the amino acid shown in the general font is part of the antibody framework.

In a preferred embodiment, the variable domains of the heavy and light chains have human origin, such as the heavy and light chain variable domains of the cesikumab antibody, which is SEQ ID NO: 10 (= light chain a variable domain, that is, amino acid 1 to 109 of SEQ ID NO: 10) and SEQ ID NO: 8 (= a variable domain of a heavy chain, that is, an amino acid of 1 to 127 of SEQ ID NO: 8) Show. The constant region preferably also comprises a suitable human constant region, for example as described in "Sequences of Proteins of Immunological Interest", Kabat E.A. et al., US Department of Health and Human Services, Public Health Service, National Institute of Health.

In some embodiments, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) comprises a light chain variable domain having SEQ ID NO: . In other embodiments, the IL-17 antagonist comprises a heavy chain variable domain having SEQ ID NO:8. In other embodiments, the IL-17 antagonist comprises a light chain variable domain of SEQ ID NO: 10 and a heavy chain variable domain of SEQ ID NO: 8. In some embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO: 10. In other embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO:8. In other embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO: 10 and three CDRs having SEQ ID NO: 8. The CDRs having SEQ ID NO: 8 and SEQ ID NO: 10, as defined by Chothia and Kabat, can be found in Table 2 .

In some embodiments, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab), comprises a light chain domain having SEQ ID NO: 15. In other embodiments, the IL-17 antagonist comprises a heavy chain domain having SEQ ID NO: 17. In other embodiments, the IL-17 antagonist comprises a light chain domain having SEQ ID NO: 15 and a heavy chain domain having SEQ ID NO: 17. In some embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO: 15. In other embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO: 17. In other embodiments, the IL-17 antagonist comprises three CDRs having SEQ ID NO: 15 and three CDRs having SEQ ID NO: 17. The CDRs having SEQ ID NO: 15 and SEQ ID NO: 17 as defined by Chothia and Kabat can be found in Table 2 .

The hypervariable regions can be combined with any kind of framework regions, although the preferred framework regions have human origin. Suitable framework regions are described in Kabat E. A. et al. (supra). A preferred heavy chain framework is a human heavy chain framework, such as the heavy chain framework of a cesikumab antibody. It consists, for example, of the following: FR1 (amino acid 1 to 30 of SEQ ID NO: 8), FR2 (amino acid 36 to 49 of SEQ ID NO: 8), FR3 (amino acid of SEQ ID NO: 8) 67 to 98) and FR4 (amino acid 117 to 127 of SEQ ID NO: 8) regions. In view of the high concentration region of the cesikumab determined by X-ray analysis, another preferred heavy chain framework consists of FR1-x (amino acids 1 to 25 of SEQ ID NO: 8), FR2. -x (amino acids 36 to 49 of SEQ ID NO: 8), FR3-x (amino acids 61 to 95 of SEQ ID NO: 8) and FR4 (amino acids 119 to 127 of SEQ ID NO: 8) Area. In a similar manner, the light chain framework consists in turn of FR1' (amino acids 1 to 23 of SEQ ID NO: 10), FR2' (amino acids 36 to 50 of SEQ ID NO: 10), FR3' (SEQ ID NO: 10 amino acid 58 to 89) and FR4' (amino acid 99 to 109 of SEQ ID NO: 10) region.

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab), is selected from a human anti-IL-17 antibody comprising at least: a) an immunoglobulin heavy chain or fragment thereof comprising a variable domain of the hypervariable regions CDR1, CDR2 and CDR3 and a constant portion of a human heavy chain or a fragment thereof; the CDR1 having the amino acid sequence SEQ ID NO: 1, The CDR2 has the amino acid sequence SEQ ID NO: 2, and the CDR3 has the amino acid sequence SEQ ID NO: 3; and b) comprises a variable domain comprising the hypervariable region CDR1', CDR2' and CDR3', and human An immunoglobulin light chain of a constant portion of a light chain or a fragment thereof, the CDR1' having an amino acid sequence of SEQ ID NO: 4, the CDR2' having an amino acid sequence of SEQ ID NO: 5, and the CDR3' It has an amino acid sequence of SEQ ID NO: 6.

In one embodiment, an IL-17 antagonist, eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), is selected from a single-stranded binding molecule comprising an antigen binding site, The antigen binding site comprises: a) a first domain comprising CDR1, CDR2 and CDR3 of the hypervariable region, the CDR1 having the amino acid sequence SEQ ID NO: 1, the CDR2 having the amino acid sequence SEQ ID NO: 2, And the CDR3 has the amino acid sequence SEQ ID NO: 3; and b) the second domain comprising the hypervariable region CDR1', CDR2' and CDR3' having the amino acid sequence SEQ ID NO: 4, the CDR2 'having an amino acid sequence of SEQ ID NO: 5, and the CDR3' has an amino acid sequence of SEQ ID NO: 6; and c) binds to the N-terminal end of the first domain and binds to the C-terminal end of the second domain or A peptide linker that binds to the C-terminal end of the first domain and binds to the N-terminal end of the second domain.

Alternatively, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab), can comprise at least one antigen binding site, the at least one antigen binding site comprising The at least one immunoglobulin heavy chain variable domain ( _VH ) is sequentially included: a) hypervariable region CDR1 (SEQ ID NO: 1), CDR2 (SEQ ID NO: 2), and CDR3 (SEQ ID NO: 3) Or b) hypervariable region CDR1 _i , CDR2 _i , CDR3 _i , the hypervariable region CDR1 _i and the hypervariable region CDR1 as shown in SEQ ID NO: 1 have 3, preferably 2, more preferably 1 amine The basal acid is different; the hypervariable region CDR2 _i and the hypervariable region CDR2 as shown in SEQ ID NO: 2 have 3, preferably 2, more preferably 1 amino acid; and the hypervariable region CDR3 _i and The high variable region CDR3 as shown in SEQ ID NO: 3 has 3, preferably 2, more preferably 1 amino acid; and the binding IL-17 antagonist can be below 50 nM or 50 nM, A concentration of about 20 nM or less, about 10 nM or less, about 5 nM or less, about 2 nM or less, or more preferably about 1 nM or less, about 1 nM or less 1 nM (=30 ng/ml) inhibits the activity of human IL-17 by 50%, and the inhibitory activity is directed against The IL-6 production induced by hu-IL-17 in human dermal fibroblasts was measured.

Likewise, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab), can comprise at least one antigen binding site, the at least one antigen binding site comprising Including at least one of the following immunoglobulin heavy chain variable domains ( _VH ): a) hypervariable regions CDR1-x (SEQ ID NO: 11), CDR2-x (SEQ ID NO: 12), and CDR3-x ( SEQ ID NO: 13); or b) hypervariable region CDR1 _i -x, CDR2 _i -x, CDR3 _i -x, the hypervariable region CDR1 _i -x and the hypervariable region CDR1 as set forth in SEQ ID NO: -x has 3, preferably 2, more preferably 1 amino acid; the hypervariable region CDR2 _i -x and the hypervariable region CDR2-x as shown in SEQ ID NO: 12 have 3, preferably 2, more preferably 1 amino acid is different; and the hypervariable region CDR3 _i -x and the hypervariable region CDR3-x as shown in SEQ ID NO: 13 have 3, preferably 2, more preferably 1 The amino acid is different; and the binding IL-17 antagonist can be at about 50 nM or less, about 20 nM or less, about 10 nM or less, about 5 nM or less, about 2 Inhibition of 1 nM (=30 ng/ml) of human IL-17 by 50% at a concentration of the molecule below nM or 2 nM, or more preferably about 1 nM or less. This inhibitory activity was measured against IL-6 production induced by hu-IL-17 in human dermal fibroblasts.

Likewise, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab), can comprise at least one antigen binding site, the at least one antigen binding site comprising the sequence comprising at least one immunoglobulin light chain variable domain (V _L): a) hypervariable regions CDR'1 (SEQ ID NO: 4) , CDR'2 (SEQ ID NO: 5) and CDR'3 ( SEQ ID NO:. 6); or b) hypervariable regions _{CDR1 'i, CDR2' i,} CDR3 'i, the hypervariable regions CDR'1 _i and as SEQ ID NO: the hypervariable regions shown with a 4 CDR'1 3, preferably 2, more preferably 1 amino acid; the hypervariable region CDR'2 _i and the hypervariable region CDR'2 as shown in SEQ ID NO: 5 have 3, preferably 2, More preferably, one amino acid is different; and the hypervariable region CDR'3 _i has three, preferably two, more preferably one amino acid as the hypervariable region CDR'3 as shown in SEQ ID NO: Different; and the binding IL-17 antagonist can be below about 50 nM or 50 nM, about 20 nM or 20 nM, about 10 nM or less, about 5 nM or less, about 2 nM or 2 The activity of 1 nM (=30 ng/ml) of human IL-17 is inhibited by 50% at a concentration of the molecule below nM, or more preferably about 1 nM or less, and the inhibitory activity is The measurement was performed against IL-6 production induced by hu-IL-17 in human dermal fibroblasts.

Alternatively, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), can comprise a heavy chain variable domain ( _VH ) and a light chain variable domain (V _L ), and the IL-17 binding molecule has at least one antigen binding site, the at least one antigen binding site comprising: a) an immunoglobulin heavy chain variable domain ( _VH ), which in turn comprises a hypervariable region CDR1 (SEQ ID NO: 1) , CDR2 (SEQ ID NO: 2) and CDR3 (SEQ ID NO: 3) ; and an immunoglobulin light chain variable domain (V _L), which in turn contains the hypervariable regions CDR1 '( SEQ ID NO: 4), CDR2' (SEQ ID NO: 5) and CDR3' (SEQ ID NO: 6); or b) immunoglobulin heavy chain comprising the hypervariable region CDR1 _i , CDR2 _i and CDR3 _i in turn a variable domain (V _H ) having 3, preferably 2, more preferably 1 amino acid, CDR1 _i as shown in SEQ ID NO: 1, the hypervariable region CDR2 _{i is} different from the hypervariable region CDR2 as shown in SEQ ID NO: 2, preferably 2, more preferably 1 amino acid, and the hypervariable region CDR3 _i is as shown in SEQ ID NO: 3. The hypervariable region CDR3 has 3, preferably 2, more preferably 1 amino acid; and sequentially comprises the hypervariable region CDR1' _i , CDR2' _i , the immunoglobulin light chain variable domain (V _L ) of CDR3' _i , the high variable region CDR ' _{1 i} and the hypervariable region CDR ' ₁ as shown in SEQ ID NO: 4 have 3, preferably 2 More preferably, the amino acid is different, and the hypervariable region CDR'2 _i and the hypervariable region CDR'2 as shown in SEQ ID NO: 5 have 3, preferably 2, more preferably 1 amino acid. Different, and the hypervariable region CDR'3 _i and the hypervariable region CDR'3 as shown in SEQ ID NO: 6 have 3, preferably 2, more preferably 1 amino acid; and the IL-17 The antagonist can be at about 50 nM or 50 nM, about 20 nM or 20 nM or less, about 10 nM or less, less than 5 nM or less, about 2 nM or less, or about 1 nM. Or inhibiting the activity of 1 nM (=30 ng/ml) human IL-17 by 50% at a concentration of 1 nM or less, which is induced by hu-IL-17 in human dermal fibroblasts. IL-6 production is measured.

Alternatively, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab), can comprise a heavy chain variable domain ( _VH ) and a light chain variable domain (V _L), and the IL-17 binding molecule comprises at least one antigen binding site, the at least one antigen binding site comprising: a) an immunoglobulin heavy chain variable domain (V _H), which in turn contains hypervariable regions CDR1-x (SEQ ID NO: 11), CDR2-x (SEQ ID NO: 12) and CDR3-x (SEQ ID NO: 13); and an immunoglobulin light chain variable domain (V _L), which in turn comprises Hypervariable regions CDR1' (SEQ ID NO: 4), CDR2' (SEQ ID NO: 5) and CDR3' (SEQ ID NO: 6); or b) sequentially comprise hypervariable regions CDR1 _i -x, CDR2 _i -x And an immunoglobulin heavy chain variable domain ( _VH ) of CDR3 _i -x, wherein the hypervariable region CDR1 _i -x and the hypervariable region CDR1-x as shown in SEQ ID NO: 11 have 3, preferably 2 More preferably, the amino acid is different, and the hypervariable region CDR2 _i -x and the hypervariable region CDR2-x as shown in SEQ ID NO: 12 have 3, preferably 2, more preferably 1 amine group. different acid, and the hypervariable region CDR3 _i -x and as SEQ ID NO: 13 shown in the hypervariable region CDR3-x is 3, preferably 2, more preferably 1 different amino acids; and by Comprising the hypervariable regions _{CDR1 'i, CDR2' i,} CDR3 ' _i of immune immunoglobulin light chain variable domain (V _L), the hypervariable regions CDR'1 _i and as SEQ ID NO: 4 of the hypervariable regions as shown in CDR'1 has 3, preferably 2, more preferably 1 amino acid, and the hypervariable region CDR'2 _i has 3 CDR '2 as shown in SEQ ID NO: 5, compared with Preferably, the two amino acids are different, and the high variable region CDR '3 _i and the high variable region CDR '3 as shown in SEQ ID NO: 6 have three, preferably two, and more preferably The amino acid is different; and the IL-17 antagonist can be below about 50 nM or 50 nM, about 20 nM or 20 nM, about 10 nM or less, about 5 nM or less, about 2 nM The activity of 1 nM (=30 ng/ml) of human IL-17 is inhibited by 50% at a concentration of the molecule of 2 nM or less, or more preferably about 1 nM or less, which is directed against hu-IL- 17 Measurement of IL-6 production induced in human dermal fibroblasts.

The human IL-17 antibody disclosed herein may comprise a heavy chain substantially identical to the heavy chain set forth as SEQ ID NO: 17 and a light chain substantially identical to the light chain set forth as SEQ ID NO: 15. The human IL-17 antibody disclosed herein may comprise a heavy chain comprising SEQ ID NO: 17 and a light chain comprising SEQ ID NO: 15. The human IL-17 antibody disclosed herein may comprise: a) a heavy chain comprising an amino acid sequence substantially identical to the variable domain of the amino acid sequence set forth in SEQ ID NO: 8 and a constant portion of the human heavy chain And b) a light chain comprising an amino acid sequence substantially identical to the variable domain of the amino acid sequence set forth in SEQ ID NO: 10 and a constant portion of the human light chain.

Inhibition of binding of IL-17 to its receptor is preferably tested in a variety of assays, including those described in WO 2006/013107. The term "to the same extent" means that the reference molecule and the derivative molecule exhibit substantially the same IL-17 inhibitory activity on a statistical basis in one of the assays referred to herein (see Example 1 of WO 2006/013107). For example, when assayed as described in Example 1 of WO 2006/013107, the IL-17 binding molecules disclosed herein inhibit human IL-17 in human IL-17-induced IL-6 production by human dermal fibroblasts. IC ₅₀ is typically less than about 10 nM, more preferably from about 9,8,7,6,5,4,3,2 or about 1 nM, preferably substantially the same as the corresponding reference molecule of the IC _50. Alternatively, the assay used can be an assay for the inhibition of binding of IL-17 by a soluble IL-17 receptor (e.g., the human IL-17 R/Fc construct of Example 1 of WO 2006/013107) in competition with an IL-17 antagonist of the invention.

The invention also includes one or more amino acid residues of CDR1, CDR2, CDR3, CDR1-x, CDR2-x, CDR3-x, CDR1', CDR2' or CDR3' or framework, usually only a few (eg 1 Up to 4 amino acid residues, such as IL-17 antagonists, such as IL-17 antibodies or antigen-binding fragments thereof, which are altered by mutation of the corresponding DNA sequence (eg, induced by site-directed mutagenesis) , for example, Sekukinumab). The invention includes DNA sequences encoding such altered IL-17 antagonists. In particular, the invention encompasses having one or more residues of CDR1' or CDR2' with the residues shown in SEQ ID NO: 4 (for CDR1') and SEQ ID NO: 5 (for CDR2') A variable IL-17 binding molecule.

The present invention also encompasses IL-17 antagonists having binding specificity for human IL-17, such as IL-17 binding molecules (e.g., IL-17 antibodies or antigen-binding fragments thereof, such as cesikumab), including, inter alia, inhibition An IL-17 antibody that binds IL-17 to its receptor, and an IL-17 antibody that is capable of about 50 nM or less, about 20 nM or less, about 10 nM or less, about 5 n. Inhibition of 1 nM (= 30 ng/ml) of human IL-17 by 50% at a concentration of the molecule of nM or 5 nM or less, about 2 nM or less, or more preferably about 1 nM or less. Inhibitory activity is measured against IL-6 production induced by hu-IL-17 in human dermal fibroblasts).

In some embodiments, an IL-17 antagonist (eg, an IL-17 antibody, eg, cesikumab) binds to mature human IL-17 and comprises the following epitopes: Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129. In some embodiments, an IL-17 antibody (eg, cesikumab) binds to mature human IL-17 comprising the following epitopes: Tyr43, Tyr44, Arg46, Ala79, Asp80. In some embodiments, an IL-17 antibody (eg, cesikumab) binds to an epitope of an IL-17 homodimer having two mature human IL-17 chains, the epitope being on a strand It contains Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129, and contains Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other chain. The residue numbering scheme used to define such epitopes is based on the following: Residue 1 is the first amino acid of the mature protein (ie, IL-17A lacks an N-terminal signal peptide with 23 amino acids and Glycine starts). The sequence of immature IL-17A is described in Swiss-Prot entry Q16552. In some embodiments, K _D IL-17 antibody is from about 100 pM to 200 pM. In some embodiments, IL-17 antibody in vitro in about 0.67 nM IC of human IL-17A bioactivity of about ₅₀ 0.4 nM. In some embodiments, the absolute bioavailability of the subcutaneous (sc) administered IL-17 antibody is in the range of from about 60% to about 80%, such as about 76%. In some embodiments, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 receptor antibody) The elimination half-life is about 4 weeks (e.g., about 23 days to about 30 days, such as about 30 days). In some embodiments, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 receptor antibody) The T _max is about 7 to 8 days.

Particularly preferred for use in IL-17 antagonists of the disclosed methods, uses, kits and the like, such as IL-17 binding molecules (eg, IL-17 antibodies or antigen-binding fragments thereof, such as cesikumab) or IL The -17 receptor binding molecule (e.g., an IL-17 antibody or antigen-binding fragment thereof) is a human antibody, particularly a cesikumab as described in Examples 1 and 2 of WO 2006/013107. Securkinumab is a recombinant high-affinity, fully human monoclonal anti-human interleukin-17A (IL-17A, IL-17) antibody of IgG1/κ isotype, which is currently in clinical treatment of immune-mediated inflammatory conditions In the test. Secukinumab plug library (see e.g. WO 2006/013107 and WO 2007/117749) it has a very high affinity for IL-17, i.e., K _D of about 100 pM to 200 pM, about 0.67 nM human and its in vitro IC IL-17A bioactivity of about ₅₀ to 0.4 nM. Thus, cesikumab inhibits the antigen at a molar ratio of about 1:1. This high binding affinity makes cesumumab antibodies particularly useful for therapeutic applications. In addition, it has been determined that cesikumab has a very long half-life, ie about 4 weeks, which allows for an extended period of time between administrations, in the treatment of chronic lifelong conditions (such as rheumatoid arthritis (RA)). Time is a superior feature.

Prediction method, diagnosis method and method for generating form of transmittable information

The disclosed method is suitable for treating, preventing or ameliorating inflammatory arthritis (e.g., rheumatoid arthritis (RA), spondyloarthropathy, ankylosing spondylitis, and dry arthritis).

The dual gene in the dual genome of the SE, HLA-DRB1*04 dual genome or the HLA-DRB1*SE dual genome can be analyzed by any conventional means (such as Western blot, northern blot, ELISA, etc.). Biological sample of the patient. The present invention is not limited by the type of analysis used to assess the presence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome in a biological sample from a patient. In fact, any well-known assay that can be used to determine the genotype status of a patient can be used to achieve the objectives of the present invention.

The invention is also not limited by the source of the biological sample, as multiple biological samples can be used to identify the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome in the patient, for example Blood, synovial fluid, white blood cell layer, serum, plasma, lymph, urine, tears, saliva, cerebrospinal fluid, buccal mucosal swab, sputum or tissue. The invention is also not limited by the source within the biological sample for identifying the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome. It will be appreciated that genomic DNA obtained from biological samples can be analyzed to detect dual genes in the SE, HLA-DRB1*04 dual genome or HLA-DRB1*SE dual genome, or HLA can be analyzed from biological samples. -DRB*04 dual gene and/or products of HLA-DRB*SE dual gene, such as nucleic acid products (such as mRNA, microRNA, etc.) and polypeptide products (such as serological antigen) to detect SE, HLA-DRB1*04 dual A dual gene in the genome or a dual gene in the HLA-DRB1*SE dual genome. In addition, the dual gene in the SE, HLA-DRB1*04 dual genome, or the dual gene in the HLA-DRB1*SE dual genome can also be detected by detecting the HLA-DRB*04 dual gene or the HLA-DRB*SE dual gene. An equivalent genetic marker can be used to determine, for example, a SNP (single nucleotide polymorphism), a microsatellite marker, another HLA dual gene (eg, an HLA-DQB1 dual gene), or other species Genetic polymorphism. For example, the presence of a genetic marker on the same simplex as the HLA-DRB1*04 dual gene rather than the HLA-DRB1*04 dual gene itself may indicate the likelihood that the patient will respond to IL-17 binding molecule therapy. A discussion of recombination and linkage disequilibrium in Class II HLA regions is provided in Begovich et al. (1992) J. Immunology 148: 249-58. Thus, the presence of an equivalent genetic marker can also be used in the disclosed methods. Preferably, the applicable equivalent genetic marker is about 200 kb or less in the relevant HLA dual gene. More preferably, the equivalent genetic marker is 100 kb or less in the relevant HLA dual gene. An exemplary equivalent genetic marker of the HLA simple type includes a SNP that marks a simple type, or a dual gene encoded by a simple type of gene (for example, a DQB1*03:02 encoding a DQ8 antigen, which is a DR4-DQ8 simple type) portion).

A variety of methods and devices for identifying the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome are well known to those skilled in the art. For example, the presence or absence of a HLA-DRB1*04 and/or HLA-DRB1*SE dual gene can be determined by directly detecting a dual gene (eg, by labeling) or by detecting a particular region therein. DNA for dual gene detection can be prepared from biological samples by methods well known in the art, such as PUREGENE DNA from Gentra Systems (Qiagen, CA). Purification system. Detecting a genomic sequence can include studying nucleotides located on a sense strand or an antisense strand in the region.

The presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome in a patient can be detected by PCR, using genomic DNA obtained by PCR, using SE, HLA-DRB1* Detection of a sequence-specific probe in a dual gene in the dual genome or a dual gene in the HLA-DRB1*SE dual genome, such as a hydrolyzed probe from Taqman, Beacons, Scorpions; or a hybridization probe . For detection, sequence-specific probes are designed to specifically hybridize to genomic DNA of the relevant HLA dual gene. Such probes can be labeled for direct detection or can be contacted with a second detectable molecule that specifically binds to the probe. PCR products can also be detected by DNA binding agents. These PCR products can then be sequenced by any of the DNA sequencing methods available in the art.

Alternatively, the presence or absence of an HLA dual gene can be detected by sequencing using any sequencing method such as, but not limited to, Sanger-based sequencing. , pyrosequencing or next generation sequencing (Shendure J. and Ji, H., Nature Biotechnology (1998), Vol. 26, No. 10, pp. 1135-1145) .

In some embodiments, hybridization assays are used to detect the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome, or a dual gene in the HLA-DRB1*SE dual genome in a patient. In hybridization assays, the presence or absence of a genetic marker is determined based on the ability of a nucleic acid in a sample to hybridize to a complementary nucleic acid molecule (eg, an oligonucleotide probe). A variety of hybridization assays are available. In some hybridization assays, hybridization of the probe to the relevant sequence is detected directly by visualizing the bound probe, such as Northern or Southern analysis. In these analyses, DNA (Southern analysis) or RNA (Northern analysis) is isolated. The DNA or RNA is then cleaved with a series of restriction enzymes that occasionally cleave in the genome and are not close to any of the markers analyzed. The DNA or RNA is then separated, for example, by agarose gel and transferred to the membrane. By, for example, incorporating a radioactive nucleotide or binding agent (eg, SYBR) Green) and the labeled probe is contacted with the membrane under conditions of low stringency, medium stringency or high stringency. Unbound probes are removed and the presence of binding is detected by visualizing the labeled probes.

Various methods of analyzing, detecting, measuring, identifying and/or determining HLA dual gene and dual gene regions are known in the art. HLA typing can be performed at low resolution, medium resolution or high resolution. Low-resolution HLA typing refers to the reporting of a dual gene at a two-digit level (eg, HLA-DRB1*04). Even if the type has been faced with the patient at the four-digit level, moderate-resolution HLA typing is also performed in the presence of ambiguity. For example, there may be only a list of known possibilities, ie HLA-DRB1*03:01-HLA-DRB1*30:01 or HLA-DRB1*03:20-HLA-DRB1*30:01 or HLA-DRB1 *03:26-HLA-DRB1*30:01. These medium resolution types can be generated by typing-based sequence-specific PCR (SSP), where a set of initial PCR primers is used to obtain a list of possible genotypes that a particular person may have (others may need to be used before a clear type is reached) The combination of the dual gene-specific primers is further tested and/or colonized and the pure lines are sequenced). However, depending on the clinical and/or research objectives of the HLA typing, additional laboratory testing may achieve a height (ie, four digits) resolution.

Low-resolution HLA typing can be achieved by antibody-based serological testing. Higher resolution can be achieved by molecular methods (DNA based methods). Such HLA typing methods include, for example, DNA amplification techniques (such as PCR and variants thereof), direct sequencing, and Luminex xMAP Technically combined sequence-specific oligonucleotide (SSO) hybridization, sequence-specific primer (SSP) typing, sequence-based typing (SBT).

Sequence-specific oligonucleotide (SSO) typing uses PCR target amplification, hybridization of the PCR product to a sequence-specific oligonucleotide set immobilized on the beads, and generation of a color-detecting binding probe amplification product. And then conduct data analysis. Those skilled in the art will appreciate that the sequence-specific oligonucleotide (SSO) hybridization can be performed using a variety of commercially available kits, such as those provided by One Lambda, Inc. (Canoga Park, CA) or With Luminex Technology (Luminex, Corporation, TX) in conjunction with the Lifecodes HLA Typing Kit (Tepnel Life Sciences Corp.). LABType SSO is a reverse SSO (rSSO) DNA typing method that recognizes HLA dual genes using sequence-specific oligonucleotide (SSO) probes and standard-labeled microspheres. The target DNA is amplified by polymerase chain reaction (PCR) and then hybridized to the bead probe array. Analysis was performed in a single well of a 96-well PCR disk; therefore, 96 samples could be processed at a time.

Sequence-specific primer (SSP) typing is a PCR-based technique using sequence-specific primers for DNA-based HLA typing. The SSP method is based on the principle that only primers having sequences that exactly match the target sequence can produce amplification products under controlled PCR conditions. A dual gene sequence-specific primer pair is designed to selectively amplify a target sequence unique to a single dual gene or a dual genome. The PCR product can be visualized on an agarose gel. Control primer pairs that match the non-dual gene sequences present in all samples serve as internal PCR controls to determine the efficiency of PCR amplification. It should be appreciated by those skilled in the art, with the low resolution sequence-specific primer typing, medium-resolution and high-resolution genotyping can be used to achieve a variety of commercially available kits, such as Olerup SSP ^TM kit (Olerup , PA) or (Invitrogen), or Allset and ^TM Gold DQA1 low resolution SSP (Invitrogen).

Sequence-based typing (SBT) is based on PCR target amplification, followed by sequencing of the PCR products and data analysis.

Messenger RNA (mRNA) levels can also be used as predictive markers to determine whether an individual is likely to respond to IL-17 binding molecule therapy. The amount of mRNA is measured using a variety of techniques known to those skilled in the art, including but not limited to Northern blot analysis, nuclease protection assay (NPA), in situ hybridization. , Reverse transcription polymerase chain reaction (RT-PCR), RT-PCR ELISA, TaqMan-based quantitative RT-PCR (probe-based quantitative RT-PCR) and SYBR Green based quantitative RT-PCR.

In one example, detecting mRNA levels involves contacting the isolated mRNA with an oligonucleotide that hybridizes to mRNA encoded by the HLA-DRB1*04 and/or HLA-DRB1*SE dual gene. A nucleic acid probe can generally be, for example, a full length cDNA or a portion thereof, such as an oligonucleotide that is at least 7, 15, 30, 50 or 100 nucleotides in length and is sufficient to specifically hybridize to mRNA under stringent conditions. Hybridization of the mRNA with the probe indicates that the marker is being expressed.

In one form, the mRNA is immobilized on a solid surface and brought into contact with the probe, for example by electrophoresing the isolated mRNA on an agarose gel and transferring the mRNA from the gel to a membrane (such as nitrocellulose). .

In another example, the mRNA content can be determined by reverse transcription polymerase chain reaction (RT-PCR), RT-PCR ELISA, TaqMan-based quantitative RT-PCR (probe-based quantitative RT-PCR), and based on SYBR green. Quantitative RT-PCR was used for the determination. Such detection protocols are particularly useful for detecting such molecules in the presence of very low numbers of nucleic acid molecules. Amplification primers are defined as a pair of nucleic acid molecules that bind to the 5' or 3' region of the gene (positive and negative strands, respectively, or vice versa) and contain short regions in between. In general, amplification primers are about 10 to 30 nucleotides in length and have a flanking region of about 50 to 200 nucleotides in length. The primers amplify a nucleic acid molecule comprising a nucleotide sequence flanked by the primers under appropriate conditions and using appropriate reagents. The PCR product can be detected by any suitable method, including, but not limited to, gel electrophoresis and staining with a DNA-specific stain or hybridization to a labeled probe.

In one embodiment, the presence of a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome in a patient is performed by using, for example, PCR-based analysis or reverse transcriptase PCR ( RT-PCR) was determined by measuring RNA content. In another aspect, quantitative RT-PCR using a standardized mixture of competing templates can be utilized.

Those skilled in the art will appreciate that analysis of HLA-DRB*04 and/or HLA-DRB*SE dual genes can be performed separately or simultaneously in the analysis of other genetic sequences. In another aspect of the invention, an array is provided, in which probes corresponding in turn to gene products (e.g., cDNA, mRNA, cRNA, polypeptide, and fragments thereof) can specifically hybridize or bind to the array at known positions.

In some embodiments, the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome is determined by measuring the polypeptide product of the HLA-DRB*04 and/or HLA-DRB*SE dual gene or in the patient. A dual gene in the HLA-DRB1*SE dual genome. The polypeptide products of HLA-DRB1*04 and/or HLA-DRB1*SE dual genes (HLA-DR4 and HLA-DRSE serological antigens) can be detected using any method known in the art, and the known methods include However, it is not limited to immunocytochemical staining, ELISA, flow cytometry, Western blotting, spectrophotometry, HPLC, and mass spectrometry. In some embodiments, serum-specific HLA typing uses antigen-specific serum to determine the patient's HLA type.

A method of detecting a polypeptide product of a HLA-DRB1*04 and/or HLA-DRB1*SE dual gene in a sample is by means of a probe which is a binding protein capable of specifically interacting with a marker protein. Preferably, labeled antibodies, binding moieties thereof, or other binding partners can be used. The origin of the antibody may be single or multiple antibodies, or the antibody may be produced biosynthetically. The binding partner can also be a naturally occurring molecule or produced synthetically. The amount of complexed protein is determined using standard protein detection methods as described in the art. A detailed review of immunological analysis designs, theories, and protocols can be found in many of the works in this technology, including Practical Immunology, Butt, W. R. ed., Marcel Dekker, New York, 1984.

A variety of assays can be used to detect proteins with labeled antibodies. Direct labeling includes fluorescent or luminescent labels attached to antibodies, metals, dyes, radionucleides, and the like. Indirect labels include various enzymes well known in the art, such as alkaline phosphatase, catalase, and the like. In a single step assay, the HLA-DRB1*04 and/or HLA-DRB1*SE dual gene polypeptide product that may be present is immobilized and incubated with the labeled antibody. The labeled antibody binds to the immobilized target molecule. After washing to remove unbound molecules, the presence of the label in the sample is analyzed.

The invention also contemplates the use of immobilized antibodies specific for a protein or polypeptide. The antibodies can be immobilized on a variety of solid supports, such as magnetic or chromatography matrix particles, surfaces of analytical sites (such as microtiter wells), solid matrix materials (such as plastic, nylon, paper) sheets, and the like. The analytical strip can be prepared by coating an antibody or a plurality of antibodies in an array on a solid support. This strip can then be immersed in the test sample and then quickly processed through the wash and detection steps to produce a measurable signal, such as a colored spot.

In a two-step assay, the immobilized HLA-DRB1*04 and/or HLA-DRB1*SE dual gene polypeptide product can be incubated with unlabeled antibodies. The unlabeled antibody complex that may be present is then bound to a second labeled antibody that is specific for the unlabeled antibody. The sample is washed and the presence of the marker is analyzed.

The choice of label for labeling antibodies will vary from application to application. However, the choice of indicia can be readily determined by those skilled in the art.

Antibodies can be labeled with radioactive atoms, enzymes, chromogenic or fluorescent moieties or colorimetric labels. The choice of marking should also be based on the required detection limits. Enzymatic analysis (ELISA) typically allows the detection of colored products formed by the interaction of an enzyme-labeled complex with an enzyme substrate. Some examples of radioactive atoms include ³² P, ¹²⁵ I, ³ H, and ¹⁴ P. Some examples of enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and glucose-6-phosphate dehydrogenase. Some examples of hair coloring moieties include luciferin and rhodamine. Antibodies can be bound to such markers by methods known in the art. For example, the enzyme and chromogenic molecule can be bound to the antibody by means of a coupling agent such as a dialdehyde, a carbodiimide, a dimethyleneimine, and the like. Alternatively, binding can be via a ligand-receptor pair. Some suitable ligand-receptor pairs include, for example, biotin-avidin or biotin-streptavidin and antibody-antigen.

In one aspect, the invention contemplates the use of sandwich techniques to detect HLA-DRB1*04 and/or HLA-DRB1*SE dual gene polypeptide products in a biological sample. This technique requires two antibodies that bind to the relevant protein: for example, one antibody is immobilized on a solid support and the other antibody is freed from solution but labeled with a compound that is readily detectable. Examples of chemical labels that can be used for the second antibody include, but are not limited to, radioisotopes, fluorescent compounds, and enzymes, or other molecules that produce colored or electrochemically active products upon exposure to the reactants or enzyme substrates. When a sample containing the HLA-DRB1*04 and/or HLA-DRB1*SE dual gene polypeptide product is placed in the system, the polypeptide product binds to both the immobilized antibody and the labeled antibody. The result is a "sandwich" immune complex on the surface of the support. The complexed protein is detected by washing away unbound sample components and excess labeled antibody and measuring the amount of labeled antibody complexed with the protein on the surface of the support. Sandwich immunoassays are highly specific and extremely sensitive, with the limitation of using markers with good detection limits.

Preferably, by radioimmunoassay or enzyme-linked immunoassay, competitive binding enzyme-linked immunoassay, spot blotting, Western blotting, chromatography (preferably high performance liquid chromatography (HPLC)) Or other assays known in the art to detect the presence of HLA-DRB1*04 and/or HLA-DRB1*SE dual gene polypeptide products in the sample. The specific immunological binding of the antibody to the protein or polypeptide can be detected directly or indirectly.

The spot blotting process is routinely performed by a skilled artisan using antibodies as probes to detect the desired protein (Promega Protocols and Applications Guide, 2nd Edition, 1991, p. 263, Promega Corporation). The sample was applied to the film using a spot blotting device. The labeled probe is incubated with the membrane and the presence of the protein is detected.

Western blot analysis is well known to the skilled artisan (Sambrook et al, Molecular Cloning, A Laboratory Manual, 1989, Vol. 3, Chapter 18, Cold Spring Harbor Laboratory). In the Western blot method, samples were separated by SDS-PAGE. Transfer the gel to the membrane. The membrane is incubated with the labeled antibody to detect the desired protein.

Cell typing can also be used for HLA typing. Representative cell analysis is mixed lymphocyte culture (MLC), which is used to determine class II HLA types. Cell analysis is more sensitive than serotyping in detecting HLA differences. This is because the small differences that are not recognized by the same antisera may stimulate T cells. This type is designated as Dw type. The serotyped DR4 was defined cytologically as DR4 Dw4, Dw10, Dw13, Dw14, Dw15.

A review of the various methods for performing HLA typing can be found in Howell et al. (2009) J Clin Pathol 2010 63: 387-390. Kits for HLA typing are available, for example, from Biotest AG, Dreiech, German; Qiagen GmbH, Germany; One Lambda Inc., Canoga Park, CA; Tepnel Corp., Stamford, CT; Olerup, PA; Luminex Corporation, Austin , TX; Abbot Molecular, IL, etc.

The analysis described above involves the following steps such as, but not limited to, immunoblotting, immunodiffusion, immunoelectrophoresis or immunoprecipitation. In some embodiments, an automated analyzer (eg, a PCR machine or an automated sequencer) is used to determine the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome and/or HLA-DRB1*SE dual genome.

Typically, the physician or genetic counselor or patient or other investigator can be informed of the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome and/or HLA-DRB1*SE dual genome. In particular, the results can be formulated into a form of conveyable information that can be communicated or transmitted to other researchers or physicians or genetic counselors or patients. This form can vary and can be tangible or intangible. The results of the presence or absence of the SE, HLA-DRB1*04 dual genome and/or the dual gene in the HLA-DRB1*SE dual genome of the individual tested may be embodied as descriptive sentences, schemas, photographs, charts, images or Any other visible form. For example, an image of a PCR product gel electrophoresis can be used to elucidate the results. The pattern showing the presence of variants in the HLA-DRB1*04 dual genome and/or HLA-DRB1*SE dual genome of an individual also applies to the indicated test results. Statements and visible forms can be recorded on tangible media, such as paper, computer readable media, such as floppy disks, compact discs, etc., or recorded on intangible media, such as e-mail or websites on the Internet or intranet. Electronic media. In addition, the results regarding the presence or absence of the SE, HLA-DRB1*04 dual genome and/or the dual gene in the HLA-DRB1*SE dual genome for the individual tested may also be recorded in sound form and transmitted via any suitable media, for example Analog or digital cable, fiber optic cable, etc., via telephone, fax, wireless mobile phone, internet telephony, and the like. All such forms (tangible and intangible) will constitute "transportable information forms". Therefore, information and information about test results can be generated anywhere in the world and transmitted to different locations. For example, when genotyping analysis is performed overseas, information and information about the test results can be generated and formulated into a transferable form as described above. Test results in a transferable form can therefore be entered into the United States. Thus, the present invention also contemplates methods for generating a form of transportable information about the presence or absence of a dual gene in the SE, HLA-DRB1*04 dual genome and/or HLA-DRB1*SE dual genome in an individual. This form of information is useful for predicting the responsiveness of patients treated with RA to treatment with an IL-17 antagonist.

Disclosed herein that a patient predicted to have rheumatoid arthritis (RA) will be treated with an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, eg, cesikumab) or An IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof) is a method of treating a likelihood of a response comprising analyzing at least one duality of the HLA-DRB1*04 dual genome in a biological sample from a patient The presence or absence of a gene, wherein the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will be against IL-17 Antagonist treatment is less likely to respond.

It is disclosed herein that a patient predicted to have RA will be an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)) A method of treating a likelihood of a response comprising: a) obtaining a biological sample from the patient; and b) analyzing the HLA-DRB1*04 dual genome in the biological sample The presence or absence of at least one dual gene, wherein the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will There is a reduced likelihood of response to treatment with an IL-17 antagonist.

It is disclosed herein that a patient predicted to have RA will be an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)) A method of treating a likelihood of a response comprising analyzing the presence or absence of a consensus epitope (SE) in a biological sample from a patient, wherein the presence of SE indicates the patient There is an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of SE indicates that the patient will have a reduced likelihood of responding to treatment with an IL-17 antagonist.

It is disclosed herein that a patient predicted to have RA will be an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)) A method of treating a likelihood of a response comprising: a) obtaining a biological sample from the patient; and b) analyzing the presence or absence of SE in the biological sample, wherein The presence of SE indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, whereas the absence of SE indicates that the patient will have a reduced likelihood of responding to treatment with an IL-17 antagonist.

Disclosed herein is the identification of a patient having RA with an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule ( For example, an IL-17 antibody or antigen-binding fragment thereof)) a method of treating responsiveness comprising: a) analyzing a biological sample from the patient to determine the presence or absence of at least one of the HLA-DRB1*04 dual genome A dual gene; and b) if the presence of the at least one dual gene is detected in the sample, the patient is designated to be responsive to treatment with an IL-17 antagonist.

Disclosed herein is the identification of a patient having RA with an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule ( For example, an IL-17 antibody or antigen-binding fragment thereof)) a method of treating responsiveness comprising: a) analyzing a biological sample from the patient to determine the presence or absence of SE; and b) detecting in the sample By the presence of at least one dual gene, the patient is designated to respond to treatment with an IL-17 antagonist.

It is disclosed herein that a patient predicted to have RA will be an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)) A method of treating a likelihood of a response comprising analyzing an at least one dual gene of the HLA-DRB1*04 dual genome in a biological sample from the patient using an automated analyzer The presence or absence, wherein the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will be bound to an IL-17 binding molecule The likelihood of treatment responding is reduced.

It is disclosed herein that a patient predicted to have RA will be an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)) A method of treating a likelihood of a response comprising analyzing the presence or absence of SE in a biological sample from the patient using an automated analyzer, wherein the presence of SE indicates that the patient will There is an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of SE indicates that the patient will have a reduced likelihood of responding to treatment with an IL-17 antagonist.

Disclosed herein are methods of generating a form of transportable information for predicting responsiveness to treatment with an IL-17 antagonist in a patient having RA comprising: a) analyzing a HLA-DRB1*04 dual genome in a biological sample from a patient The presence or absence of at least one dual gene; and b) the actualization of the results of the analysis step into a form of transportable information.

Disclosed herein are methods of generating a form of transportable information for predicting responsiveness to treatment with an IL-17 antagonist in a patient having RA comprising: a) analyzing the presence or absence of SE in a biological sample from a patient; b) The result of the analysis step is embodied in the form of transportable information.

Also disclosed herein is an in vitro test method for selecting a patient for RA treatment comprising determining the presence or absence of at least one dual gene in the HLA-DRB1*04 dual genome, presence or absence of the HLA-DRB1*SE dual genome in the patient. At least one dual gene or SE. In some embodiments, a patient having a SE, or a dual gene in the HLA-DRB1*04 dual genome, or a dual gene in the HLA-DRB1*SE dual genome, has an improved therapeutic response to the following regimen: a) administration of the The patient received three doses of approximately 10 mg/kg IL-17 antagonist, the first dose was delivered during week 0, the second dose was delivered during week 2, and the third dose was delivered during week 4; and b) thereafter The patient is administered about 75 mg to about 300 mg of IL-17 antagonist twice a month, once a month, once every two months, or once every three months, starting during week 8.

In some embodiments of the above methods, the presence of two dual genes in the HLA-DRB1*04 dual genome in the biological sample indicates that the patient will further increase the likelihood of responding to treatment with the IL-17 antagonist. In some embodiments, the presence of two dual genes in the HLA-DRB1*SE dual genome in the biological sample indicates that the patient will further increase the likelihood of responding to treatment with the IL-17 antagonist.

In some embodiments, the presence or absence of SE is detected by analyzing the presence or absence of at least one of the dual genes in the HLA-DRB1*SE dual genome in the biological sample.

In some embodiments, the presence or absence of at least one dual gene is detected by analyzing an genomic sequence of at least one dual gene in the biological sample, a product of at least one dual gene, or an equivalent genetic marker of at least one dual gene.

In some embodiments, the presence or absence of at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or the HLA-DRB1*SE dual genome is by a population selected from the group consisting of Techniques for detection: Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR), RT-PCR ELISA, TaqMan-based quantitative RT-PCR (probe-based quantitative RT-PCR), SYBR Green based quantification RT-PCR, polymerase chain reaction (PCR), direct sequencing, sequence-specific oligonucleotide (SSO) hybridization, sequence-specific primer (SSP) typing, and sequence-based typing (SBT), southern ink Point method, quantitative PCR (probe based or based on SYBR green), immunoassay, immunohistochemistry, ELISA, flow cytometry, Western blot, HPLC and mass spectrometry.

In some embodiments, the biological sample is selected from the group consisting of synovial fluid, blood, serum, plasma, urine, tears, saliva, cerebrospinal fluid, white blood cell samples, and tissue samples.

In some embodiments, the patient is a high risk RA patient.

Therapeutic methods and use of IL-17 antagonists

The disclosed IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or Antigen-binding fragments) are indicated for the treatment, prevention or amelioration of RA (eg signs and symptoms and structural changes, prevention of further joint erosion, improvement of joint structure, etc.), in particular for the treatment, prevention or amelioration of SE, HLA-DRB1*04 dual genomes RA of a RA patient with a dual gene or a dual gene in the HLA-DRB1*SE dual genome.

An IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof) Can be used in vitro, ex vivo, or incorporated into a pharmaceutical composition and administered to an individual (eg, a human patient) in vivo to treat, ameliorate, or prevent, for example, a dual gene or HLA in the dual genome of SE, HLA-DRB1*04 RA of RA patients with a dual gene in the DRB1*SE dual genome. Pharmaceutical compositions are formulated to be compatible with their intended route of administration (e.g., oral compositions generally include an inert diluent or an edible carrier). Other non-limiting examples of routes of administration include parenteral (e.g., intravenous), intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration. Pharmaceutical compositions that are compatible with each of the intended routes are well known in the art.

An IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof) It can be used in the form of a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. The compositions may contain, in addition to the IL-17 antagonist, carriers, various diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The characteristics of the carrier will depend on the route of administration. The pharmaceutical compositions of the present invention may be in the form of a liposome wherein, in addition to other pharmaceutically acceptable carriers, the IL-17 antagonist is also combined with an amphoteric agent (such as a lipid) which acts in agglomerated form. The cell, insoluble monolayer, liquid crystal or flake layer is present in the aqueous solution. Lipids suitable for use in liposome formulations include, without limitation, monoglycerides, diglycerides, thioesters, lysolecithin, phospholipids, saponins, bile acids, and the like.

The pharmaceutical compositions used in the disclosed methods may also contain other therapeutic agents for treating a particular targeted condition. For example, the pharmaceutical composition may also include an anti-inflammatory agent. These other factors and/or agents may be included in the pharmaceutical composition to produce a synergistic effect with the IL-17 binding molecule or to minimize side effects caused by the IL-17 antagonist, such as for example An IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof).

When orally administered a therapeutically effective amount of an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, In the case of an IL-17 antibody or antigen-binding fragment thereof, the binding agent should be in the form of a troche, capsule, powder, solution or elixirs. When administered in the form of a lozenge, the pharmaceutical compositions of the present invention may additionally contain a solid carrier such as gelatin or an adjuvant. When administered in liquid form, a liquid carrier such as an oil of oil, petroleum, animal or vegetable origin (such as peanut oil (beware of peanut allergy), mineral oil, soybean oil or sesame oil) or synthetic oil may be added. The pharmaceutical composition in liquid form may further contain a component such as a physiological saline solution, dextrose or other sugar solution or a glycol such as ethylene glycol, propylene glycol or polyethylene glycol.

When administered by intravenous, cutaneous or subcutaneous injection, a therapeutically effective amount of an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or IL- In the case of a 17 receptor binding molecule (e.g., an IL-17 antibody or antigen binding fragment thereof), the IL-17 antagonist should be in the form of a non-pyrogenic, parenterally acceptable solution. Pharmaceutical compositions for intravenous, cutaneous or subcutaneous injection may contain, in addition to the IL-17 antagonist, an isotonic vehicle such as sodium chloride, Ringer's, dextrose, dextrose and Sodium chloride, lactated Ringer's reagent, or other vehicles known in the art.

Pharmaceutical compositions for use in the disclosed methods can be made in a conventional manner. In one embodiment, the pharmaceutical composition is provided in lyophilized form. For immediate administration, it is dissolved in a suitable aqueous carrier such as sterile water for injection or sterile buffered saline. If it is considered necessary to formulate a larger volume solution for administration by infusion rather than rapid injection, it is advantageous to incorporate human serum albumin or the patient's own heparinized blood into physiological saline during formulation. The presence of an excess of such physiologically inert proteins prevents loss of the antibody by adsorption to the vessel and tube wall for infusion of the solution. If albumin is used, a suitable concentration is from 0.5% by weight to 4.5% by weight of the physiological saline solution. Other formulations include liquid or lyophilized formulations.

The antibody (e.g., an antibody against IL-17) is typically formulated as a lyophilizate for parenteral administration or as a lyophilizate that is reconstituted with a suitable diluent prior to administration. In some embodiments of the disclosed methods and uses, an IL-17 antagonist, such as an IL-17 antibody, such as a cesikumab line, is formulated as a lyophilizate. Suitable lyophilized formulations can be reconstituted in small liquid volumes (e.g., 2 ml or less) to allow subcutaneous administration, and solutions containing lower levels of antibody aggregates can be provided. Antibodies are now widely used as an active ingredient of a pharmaceutical, comprising the product HERCEPTIN ^TM (trastuzumab (trastuzumab)), RITUXAN ^TM (rituximab (rituximab)), SYNAGIS ^TM (palivizumab (Palivizumab)) Wait. Techniques for purifying antibodies to pharmaceutical grade are well known in the art.

The appropriate dose will of course depend, for example, on the particular IL-17 antagonist to be used, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule ( For example, the IL-17 antibody or antigen-binding fragment thereof, the host, the mode of administration, and the nature and severity of the condition being treated, and will vary depending on the nature of the prior treatment the patient is undergoing. Ultimately, the attending health care provider will determine the amount of IL-17 antagonist to treat each individual patient. In some embodiments, the attending health care provider can administer a low dose of an IL-17 antagonist and observe the patient's response. In other embodiments, the initial dose of the IL-17 antagonist administered to the patient is higher and then titrated down until the recurrence sign appears. Large doses of IL-17 antagonist can be administered until the patient has the optimal therapeutic effect, and the dose is generally not further increased.

The IL-17 binding molecule is preferably administered parenterally, intravenously, for example, to the anterior elbow or other peripheral vein, intramuscularly or subcutaneously. The duration of intravenous (i.v.) therapy using the pharmaceutical compositions of the present invention will vary depending on the severity of the condition being treated and the condition and individual response of the individual patient. Subcutaneous (s.c.) therapies using the pharmaceutical compositions of the invention are also contemplated. The health care provider will determine the appropriate duration of i.v. or s.c. therapy using the pharmaceutical compositions of the present invention and the time course of administration of the therapy.

Preferred administration and treatment regimens for the treatment of RA patients with a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome are provided in Table 3 :

Table 3 : Preferred dosing regimens for the treatment of RA patients

The time course of administration is generally measured from the day of the first administration (also referred to as "baseline") of the active compound (eg, secukimumab). However, different health care providers use different naming conventions, as shown in Table 4 below.

Table 4 - Common nomenclature rules for dosing regimens. Bold items refer to the naming rules used in this article.

To be consistent, as used herein, the first dose will be referred to as week 0, and the first day of dosing will be referred to as day 1. Thus, for example, four initial doses of serecumab administered once a week during the induction regimen should be between week 0 (eg, on about day 1), during week 1 (eg, at Approximately 8 days), provided during the second week (eg, on approximately day 15) and during the third week (eg, on approximately day 22). It will be appreciated that it is not necessary to provide a dose at an accurate time point, for example a dose intended to be provided on day 29 may be provided, for example, from day 24 to day 34.

It will be appreciated that for certain patients, for example, an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, such as cesikumab) or an IL-17 receptor binding molecule Treatment (eg, IL-17 antibodies or antigen-binding fragments thereof) in patients who show a lack of response may require an increased dose (eg, during the induction and/or maintenance period). Thus, the subcutaneous dose of cesumumab may be greater than about 75 mg to about 300 mg (subcutaneous), such as about 80 mg, about 100 mg, about 125 mg, about 175 mg, about 200 mg, about 250 mg, about 350. Mg, about 400 mg, etc.; likewise, the intravenous dose may be greater than about 10 mg/kg, such as about 11 mg/kg, 12 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/ Kg, 35 mg/kg, etc. It should also be understood that for some patients, for example, patients who show an adverse event or adverse reaction with an IL-17 antagonist (eg, cesagumab) may also need to reduce the dose (eg during the induction and/or maintenance period). ). Thus, the dose of cesumumab may be less than about 75 mg to about 300 mg (subcutaneous), such as about 25 mg, about 50 mg, about 80 mg, about 100 mg, about 125 mg, about 175 mg, about 200 mg. 250 mg, etc. Similarly, the intravenous dose may be less than about 10 mg/kg, such as about 9 mg/kg, 8 mg/kg, 5 mg/kg, 4 mg/kg, 3 mg/kg, 2 mg/kg, 1 mg/kg, etc.

Disclosed herein are methods of treating rheumatoid arthritis (RA) comprising: a) obtaining a biological sample from a RA patient; and b) analyzing the presence of at least one dual gene in the HLA-DRB1*04 dual genome in the biological sample or Not present; and c) if at least one dual gene is present in the biological sample, the RA patient is administered a therapeutically effective amount of an IL-17 antagonist (eg, secukimumab).

Disclosed herein is a method of treating RA comprising: a) analyzing the presence or absence of at least one dual gene in a HLA-DRB1*04 dual genome in a biological sample from a RA patient; and b) if at least one of the biological sample is present For a dual gene, a therapeutically effective amount of an IL-17 antagonist (e.g., cesikumab) is administered to the RA patient.

Disclosed herein is a method of selectively treating a patient having RA comprising: a) determining the presence or absence of a biological sample from the patient in the presence or absence of at least one dual gene in the HLA-DRB1*04 dual genome; and b) if the biological sample Where at least one dual gene is present, a therapeutically effective amount of an IL-17 antagonist (e.g., secukimumab) is administered to the RA patient.

Disclosed herein are methods of treating a patient with RA comprising: a) receiving information regarding the presence or absence of at least one dual gene in the HLA-DRB1*04 dual genome in a biological sample obtained from the patient; and b) receiving the data The data indicates that the patient has at least one dual gene and is administered the patient's IL-17 antagonist (eg, cesikumab).

Disclosed herein are methods of treating RA comprising: a) obtaining a biological sample from a RA patient; and b) analyzing the presence or absence of a consensus epitope (SE) in the biological sample; and c) if an SE is present in the biological sample A therapeutically effective amount of an IL-17 antagonist (e.g., cesikumab) is administered to the RA patient.

Disclosed herein are methods of treating RA comprising: a) analyzing the presence or absence of SE in a biological sample from a RA patient; and b) administering a therapeutically effective amount of IL to the RA patient if SE is present in the biological sample. 17 antagonist (eg, cesikumab).

Disclosed herein is a method of selectively treating a patient having RA comprising: a) determining the presence or absence of SE in a biological sample from the patient; and b) administering the RA patient if SE is present in the biological sample A therapeutically effective amount of an IL-17 antagonist (eg, cesikumab).

Disclosed herein are methods of treating a patient with RA comprising: a) receiving data regarding the presence or absence of SE in a biological sample obtained from the patient; and b) if the received data indicates that the patient has at least one dual gene, then With this patient an IL-17 antagonist (eg, cesikumab).

Disclosed herein are methods of treating RA comprising: a) selecting a RA patient for treatment based on the RA patient having at least one dual gene in the HLA-DRB1*04 dual genome; and b) administering a therapeutically effective amount to the RA patient An IL-17 antagonist (eg, cesikumab).

Disclosed herein is a method of treating RA comprising administering an IL-17 antagonist (e.g., cesikumab) to a patient with RA, the RA patient having an increased likelihood of responding to the IL-17 antagonist, wherein the likelihood is increased This is determined based on the patient having at least one dual gene in the HLA-DRB1*04 dual genome.

Disclosed herein is a method of treating RA comprising administering an IL-17 antagonist (e.g., cesikumab) to a patient with RA, the RA patient having an increased likelihood of responding to the IL-17 antagonist, wherein the likelihood is increased It is identified by the prediction method in this paper.

Disclosed herein is a method of treating RA comprising administering a therapeutically effective amount of an IL-17 antagonist (e.g., cesikumab) to a patient with RA, the condition being that the RA patient has at least one duality of the HLA-DRB1*04 dual genome. gene.

Disclosed herein are methods of treating RA comprising: a) selecting a patient with RA for treatment based on the patient having RA; and b) administering a therapeutically effective amount of an IL-17 antagonist to the patient with RA (eg, Sekkin anti).

Disclosed herein is a method of treating RA comprising administering an IL-17 antagonist (e.g., cesikumab) to a patient with RA, the RA patient having an increased likelihood of responding to the IL-17 antagonist, wherein the likelihood is increased It is determined based on the patient having SE.

Disclosed herein is a method of treating RA comprising administering an IL-17 antagonist (e.g., cesikumab) to a patient with RA, the RA patient having an increased likelihood of responding to the IL-17 antagonist, wherein the likelihood is increased It is identified by any of the methods disclosed herein.

Disclosed herein is a method of treating RA comprising administering a therapeutically effective amount of an IL-17 antagonist (e.g., secukimumab) to a patient with RA, the condition being that the RA patient has SE.

In some embodiments, the administering step comprises intravenously administering to the patient three doses of about 10 mg/kg of an IL-17 antagonist (eg, cesikumab), each of which is administered once every other week. In some embodiments, the administering step comprises subcutaneously administering about 75 mg to about 300 mg of the IL-17 antagonist to the patient twice a month, once a month, once every two months, or once every three months.

Also disclosed herein are IL-17 antagonists (e.g., cesikumab) for the treatment of RA, characterized by: a) obtaining a biological sample from a RA patient; b) analyzing the HLA-DRB1*04 dual genome in the biological sample. The presence or absence of at least one dual gene; and c) administration of a therapeutically effective amount of an IL-17 antagonist to the RA patient if at least one dual gene is present in the biological sample.

Disclosed herein are IL-17 antagonists (e.g., cesikumab) for the treatment of RA, characterized by: a) analyzing the presence of at least one dual gene in the HLA-DRB1*04 dual genome in a biological sample from a RA patient Or absent; and b) if at least one dual gene is present in the biological sample, the therapeutically effective amount of the IL-17 antagonist is administered to the RA patient.

Disclosed herein are IL-17 antagonists (eg, cesikumab) for treating RA, characterized by: a) obtaining a biological sample from a RA patient; b) analyzing the presence or absence of SE in the biological sample; If at least one dual gene is present in the biological sample, a therapeutically effective amount of an IL-17 antagonist is administered to the RA patient.

Disclosed herein are IL-17 antagonists (eg, cesikumab) for treating RA, characterized by: a) analyzing the presence or absence of SE in a biological sample from a RA patient; and b) if the biological sample is in the sample In the presence of at least one dual gene, a therapeutically effective amount of an IL-17 antagonist is administered to the RA patient.

Disclosed herein are IL-17 antagonists (eg, cesikumab) for the treatment of RA, characterized by: a) selecting a RA patient for treatment based on the RA patient having at least one of the HLA-DRB1*04 dual genomes a dual gene; and b) administering a therapeutically effective amount of an IL-17 binding molecule to the RA patient.

Disclosed herein are IL-17 antagonists (e.g., cesikumab) for treating a patient having RA, characterized in that the IL-17 antagonist is to be administered with at least one duality of the HLA-DRB1*04 dual genome Gene patients.

Disclosed herein are IL-17 antagonists (eg, cesikumab) for treating a patient having RA, characterized in that the IL-17 antagonist is to be administered based on at least one of having a HLA-DRB1*04 dual genome A patient who has been selected for treatment with a dual gene.

Disclosed herein are IL-17 antagonists (eg, cesikumab) for the treatment of RA, characterized by: a) selecting a patient with RA for treatment based on the patient having RA; and b) administering the patient with RA A therapeutically effective amount of an IL-17 antagonist.

Disclosed herein are IL-17 antagonists (e.g., cesikumab) for treating a patient having RA, characterized in that the IL-17 antagonist is to be administered to a patient having SE.

Disclosed herein are IL-17 antagonists (e.g., secukimenumab) for treating a patient having RA, characterized in that the IL-17 antagonist is to be administered to a patient selected for treatment based on having SE.

In some embodiments, three doses of about 10 mg/kg of an IL-17 antagonist (e.g., cesikumab) are administered intravenously to a patient in need thereof, each dose being delivered once every other week. In some embodiments, the patient is administered a subcutaneous administration of an IL-17 antagonist at a dose of from about 75 mg to about 300 mg subcutaneously twice a month, once monthly, once every two months, or once every three months.

Also disclosed herein is the use of an IL-17 antagonist (e.g., secukizumab) for the manufacture of a medicament for treating a patient having RA, wherein the RA patient has at least one of the HLA-DRB1*04 dual genome Dual gene.

Disclosed herein is the use of an IL-17 antagonist (eg, cesikumab) for the manufacture of a medicament for treating a patient having RA, wherein the RA patient is based on having at least one of the HLA-DRB1*04 dual genome The dual gene is selected for treatment.

Disclosed herein is the use of an IL-17 antagonist (eg, cesikumab) for the manufacture of a medicament for treating a patient having RA, wherein the RA patient has SE.

Disclosed herein is the use of an IL-17 antagonist (eg, cesikumab) for the manufacture of a medicament for treating a patient having RA, wherein the RA patient is selected for treatment based on having SE.

Disclosed herein is the use of an IL-17 antagonist for the preparation of a medicament for the treatment of RA, the restriction being based on the dual gene in the dual genome of the SE, HLA-DRB1*04 dual genome or the HLA-DRB1*SE dual genome The dual gene is selected for treatment.

Disclosed herein is the use of an IL-17 antagonist for the manufacture of a medicament for treating RA in a patient characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome or a HLA-DRB1*SE dual genome The dual gene, wherein the drug is formulated to comprise a container, each container having a sufficient amount of an IL-17 antagonist to deliver at least about 75 mg to about 150 mg of the IL-17 antagonist per unit dose.

Disclosed herein is the use of an IL-17 antagonist for the manufacture of a medicament for treating RA in a patient characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome or a HLA-DRB1*SE dual genome The dual gene, wherein the drug is formulated to contain a container, each container having a sufficient amount of an IL-17 antagonist to deliver at least about 10 mg of IL-17 antagonist per kilogram of patient body weight per unit dose.

Disclosed herein is the use of an IL-17 antagonist for the manufacture of a medicament for treating RA in a patient characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome or a HLA-DRB1*SE dual genome The dual gene, wherein the drug is formulated at a dose such that each unit dose intravenously delivers about 10 mg of IL-17 antagonist per kilogram of patient body weight.

Disclosed herein is the use of an IL-17 antagonist for the manufacture of a medicament for treating RA in a patient characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome or a HLA-DRB1*SE dual genome The dual gene, wherein the drug is formulated at a dose such that about 75 mg to about 150 mg of the IL-17 antagonist is delivered subcutaneously per unit dose.

As used herein, the phrase "a container having a sufficient amount of an IL-17 antagonist to permit delivery of a [specified dose]" is used to mean that a predetermined container (eg, vial, pen injector, syringe) is placed to provide the desired dose. A volume of an IL-17 antagonist (eg, as part of a pharmaceutical composition). For example, if the required dose is 75 mg, the clinician can use a 2 ml formulation in a container containing a concentration of 37.5 mg/ml of the IL-17 antibody formulation, containing a concentration of 75 mg/ml of IL- 1 ml of the formulation in the container of the 17 antibody formulation, 0.5 ml of the formulation in a container containing the IL-17 antibody formulation at a concentration of 150 mg/ml, and the like. In each of these conditions, the containers have a sufficient amount of IL-17 antagonist to allow delivery of the desired 75 mg dose.

The phrase "dispensing at a dose to allow [delivery route] delivery [specified dose]" as used herein is used to mean that a given pharmaceutical composition can be used to provide a desired dose of IL via a prescribed route of administration (eg, subcutaneously or intravenously). An -17 antagonist, such as an IL-17 antibody, such as cesikumab. For example, if the required subcutaneous dose is 75 mg, the clinician can use a 2 ml IL-17 antibody formulation at a concentration of 37.5 mg/ml, a 1 ml IL-17 antibody formulation at a concentration of 75 mg/ml, 0.5 ml IL-17 antibody formulation at a concentration of 150 mg/ml, and the like. In each of these conditions, the concentration of such IL-17 antibody formulations is sufficiently high to allow subcutaneous delivery of IL-17 antibodies. Subcutaneous delivery typically requires delivery of a volume of less than about 2 ml, preferably about 1 ml or less.

In some embodiments, the presence or absence of SE is detected by analyzing the presence or absence of at least one of the dual genes in the HLA-DRB1*SE dual genome in the biological sample.

In some embodiments, the presence or absence of at least one dual gene is detected by analyzing an genomic sequence of at least one dual gene in the biological sample, a product of at least one dual gene, or an equivalent genetic marker of at least one dual gene.

In some embodiments, the presence or absence of at least one of the dual gene in the SE, HLA-DRB1*04 dual genome or the HLA-DRB1*SE dual genome is by a population selected from the group consisting of Techniques for detection: Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR), RT-PCR ELISA, TaqMan-based quantitative RT-PCR (probe-based quantitative RT-PCR), SYBR Green based quantification RT-PCR, polymerase chain reaction (PCR), direct sequencing, sequence-specific oligonucleotide (SSO) hybridization, sequence-specific primer (SSP) typing, and sequence-based typing (SBT), southern ink Point method, quantitative PCR (probe based or based on SYBR green), immunoassay, immunohistochemistry, ELISA, flow cytometry, Western blot, HPLC and mass spectrometry.

In some embodiments, the biological sample is selected from the group consisting of synovial fluid, blood, serum, plasma, urine, tears, saliva, cerebrospinal fluid, white blood cell samples, and tissue samples.

In some embodiments, the patient is a high risk RA patient.

Combination therapy for RA

In practicing the methods of treatment or use of the invention, administering to a patient (eg, a mammal (eg, a human)) a therapeutically effective amount of an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, For example, cesikumab) or an IL-17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof)). An IL-17 antagonist (eg, cesikumab) can be administered alone or in combination with other dual-genes in the dual genome of the SE, HLA-DRB1*04 dual genome or the HLA-DRB1*SE dual genome in accordance with the methods of the invention. Administration of a combination of agents and therapies of RA patients with dual genes, for example, in combination with at least one anti-rheumatic agent, such as an immunosuppressant, a disease-modifying antirheumatic drug (DMARD), a pain-control drug, a steroid, a non-steroid Anti-inflammatory drugs (NSAIDs), cytokine antagonists, bone assimilating agents, bone anti-absorbers, and combinations thereof (eg, dual and triple therapy). When co-administered with one or more other agents, the IL-17 antagonist can be administered concurrently or sequentially with other agents. If administered sequentially, the attending physician will decide on the appropriate sequence for administration of the IL-17 antagonist and other agents.

Non-steroidal anti-inflammatory drugs and pain control agents suitable for use in combination with securkinumab in the treatment of RA patients include propionic acid derivatives, acetic acid derivatives, enolic acid derivatives, fenamic acid derivatives, Cox inhibitors, such as Lu Lumiracoxib, ibuprophen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, indomethacin Indomethacin), sulindac, etodolac, ketorolac, nabumetone, aspirin, naproxen, valdecoxib ), etoricoxib, MK0966; rofecoxib, acetominophen, Celecoxib, Diclofenac, tramadol, piroxicam ( Piroxicam), meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, mefanamic acid, armor Meclofenamic acid, flufenamic acid, tofininamic, cutting Valdecoxib, parecoxib (parecoxib), etodolac, indomethacin, aspirin, ibuprofen, a non-Luo rofecoxib (firocoxib). Suitable for use in combination with an IL-17 antagonist (eg, cesomumab) for the treatment of RA patients with a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome DMARD includes methotrexate (MTX), antimalarials (such as hydroxylated chloroquine and chloroquine), sulfasalazine, leflunomide, azathioprine, cyclosporin ), gold salt, minocycline, cyclophosphamide, D-penicillamine, minocycline, auranofin, tacrolimus , sodium thiobenzoate (myocrisin), chlorambucil. Suitable for use in combination with an IL-17 antagonist (eg, cesomumab) for the treatment of RA patients with a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome Steroids (eg, glucocorticoids) include Prednisolone, Prednisone, dexamethasone, cortisol, cortisone, hydrocortisone, Methylprednisolone, betamethasone, triamcinolone, beclometasome, fludrocottisone, deoxycorticosterone, aldosterone.

Suitable for use in combination with an IL-17 antagonist (eg, cesomumab) for the treatment of RA patients with a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome The biological agent is adalimumab (ADALIMUMAB) (Humira ), etanercept (ETANERCEPT) (Enbrel) ), Infliximab (INFLIXIMAB) (Remicade) ;TA-650), Certizumab PEGOL (Cimzia) ; CDP870), golimumab (Simponi) ;CNTO148), human recombinant anakinra (ANAKINRA) (Kineret ), rituximab (Rituxan) ;MabThera ), ABABACEPT (Orencia) ), tocilizumab (TOCILIZUMAB) (RoActemra/Actemra ), integrin antagonist (TYSABRI) (Nalizumab), IL-1 antagonist (ACZ885 (Ilaris), human recombinant anakinra (Kineret) )), CD4 antagonists, other IL-17 antagonists (LY2439821, RG4934, AMG827, SCH900117, R05310074, MEDI-571, CAT-2200), IL-23 antagonists, IL-20 antagonists, IL-6 antagonists a TNFα antagonist (eg, a TNFα antagonist or a TNFα receptor antagonist, such as pegsunercept, etc.), a BLyS antagonist (eg, Atacicept, Benlysta) /LymphoStat-B (belimumab), P38 inhibitor, CD20 antagonist (Ocrelizumab, Ofatumumab) (Arzerra) )), an interferon gamma antagonist (Fontolizumab).

Other RA patients suitable for treatment with a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome in combination with an IL-17 antagonist (eg, cesumumab) Agents include SB-681323, Rob 803, AZD5672, AD 452, SMP 114, HZT-501, CP-195, 543, Doxycycline, vancomycin, CRx-102, AMG108, pioglitazone , SBI-087, SCIO-469, Cura-100, Oncoxin + Viusid, TwHF, PF-04171327, AZD567, psoralen (Methoxsalen), ARRY-438162, vitamin D-wheat Ceramide (ergocalciferol), milnacipran, paclitaxel, GW406381, rosiglitazone, SC12267 (4SC-101); LY2439821, BTT-1023, ERB-041, ERB- 041, KB003, CF101, ADL5859, MP-435, ILV-094, GSK706769, GW856553, ASK8007, MOR103, HE3286, CP-690, 550 (tasocitinib), REGN88 (SAR153191), TRU-015, BMS- 582949, SBI-087, LY2127399, E-551S-551, H-551, GSK3152314A, RWJ-445380, tacrolimus (Prograf ), RAD001, rapamune, rapamycin, fostamatinib, fentanyl, XOMA 052, CNTO 136, JNJ 38518168, Imatinib, ATN -103, ISIS 104838, folic acid, folate, TNFα cytokines, MM-093, type II collagen, VX-509, AMG 82770, masitinib (AB1010), LY2127399, cyclosporine Cyclosporine, SB-681323, MK0663, NNC 0151-0000-0000, ATN-103, CCX 354-C, CAM3001, LX3305, Cetrorelix, MDX-1342, TMI-005, MK0873, CDP870 , Tranilast, CF101, mycophenolic acid (and its esters), VX-702, GLPG0259, SB-681323, BG9924, ART621, LX3305, T-614, flumarotinid disodium ( R935788), CCI-779, ARRY-371797, CDP6038, AMG719, BMS-582949, GW856553, rosiglitazone, CH-4051, CE-224, 535, GSK1827771, GW274150, BG9924, PLX3397, TAK-783, INCB028050, LY2127399, LY3009104, R788, curcumin (curcumin) (Longvida ^TM), rosuvastatin (rosuvastatin), PRO283698, AMG 714 , MTRX1011A, Maraviroc, MEDI-522, MK0663, STA 5326 mesylate, CE-224, 535, AMG108, BG00012, ramipril, VX-702, CRx-102, LY2189102, SBI-087, SB-681323, CDP870, milnacipran, PD 0360324, PH-797804, AK106-001616, PG-760564, PLA-695, MK0812, ALD518, Cobiprostone, somatotropin, tgAAC94 Gene therapy vectors, MK0359, GW856553, esomeprazole, everolimus, trastuzumab, bone assimilation agents, and bone anti-absorbers (eg, PTH, bisphosphonates (eg, azoles) Zoledronic acid, JAK1 and JAK2 inhibitors, total JAK inhibitors such as tetracyclone 6 (P6), 325, PF-956980, osteosclerin antagonist (eg BPS804), denosumab Monoclonal antibody (denosumab), IL-6 antagonist, CD20 antagonist, CTLA4 antagonist, IL-8 antagonist, IL-21 antagonist, IL-22 antagonist, integrin antagonist (Tysarbri (natalizumab), osteosclerosis antagonist, VGEF antagonist, CXCL antagonist, MMP antagonist, defensin antagonist, IL-1 antagonist (including IL-1 beta antagonist) And IL-23 antagonists (eg, receptor decoys, antagonistic antibodies, etc.).

The skilled artisan will be able to discern the appropriate dosage of the above agents that are co-delivered with serkumumab.

Set and probe

The invention also encompasses a kit for detecting a dual gene in a SE, HLA-DRB1*04 dual genome, or a dual gene in the HLA-DRB1*SE dual genome in a biological sample (test sample) from a patient. These kits can be used to predict whether a patient with RA is likely to be an IL-17 antagonist, such as an IL-17 binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof, such as cesikumab) or IL. A -17 receptor binding molecule (eg, an IL-17 antibody or antigen-binding fragment thereof) is therapeutically responsive (or has a higher response). For example, the kit can comprise a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome, a product of the dual gene, and/or in the biological sample. Or probes of equivalent genetic markers of such dual genes (eg, oligonucleotides, antibodies, labeled compounds, or other reagents).

The probe may be an oligonucleotide or a binding oligonucleotide that specifically hybridizes to a particular region within the HLA dual gene genetic marker; a PCR primer that, along with another primer, amplifies the specificity within the HLA dual gene genetic marker a region; an antibody that recognizes an HLA dual gene genetic marker and/or a polypeptide product of the HLA dual genetic marker. Optionally, the kit may contain a probe that targets an internal control dual gene, which may be any dual gene present in the general population. The detection of internal control dual genes is designed to ensure the effectiveness of the kit. The disclosed kits may also contain, for example, buffers, preservatives or protein stabilizers. The kit can also contain the components required to detect a detectable factor (eg, an enzyme or a substrate). The kit may also contain a control sample or a series of control samples that can be analyzed and compared to the test sample contained. The components of the kit are typically contained in individual containers, and all of the containers are in a single package along with their instructions for use.

Such kits may also comprise an IL-17 antagonist (eg, an IL-17 binding molecule (eg, an IL-17 antibody or antigen binding fragment thereof, eg, cesikumab) or an IL-17 receptor binding molecule (eg, IL-) 17 antibody or antigen-binding fragment thereof)) (for example in liquid or lyophilized form) or a pharmaceutical composition comprising an IL-17 antagonist (described above). In addition, the kits can include components for administering an IL-17 antagonist (eg, a syringe or a prefilled pen injector) and instructions for use. Such kits may contain, for example, other therapeutic agents (described above) for use in the treatment of RA, which are delivered in combination with an IL-17 antagonist (e.g., secukinumab).

Disclosed herein are kits for predicting the likelihood that a patient with rheumatoid arthritis (RA) will respond to treatment with an IL-17 antagonist (eg, sequenumab) comprising: a) at least one a probe capable of detecting the presence of at least one dual gene in the HLA-DRB1*04 dual genome; and b) instructions for using the probe to analyze the presence of at least one dual gene in a biological sample from the RA patient, wherein The at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will be less likely to respond to treatment with an IL-17 antagonist. .

Disclosed herein are kits for treating a patient having RA comprising: a) a therapeutically effective amount of an IL-17 antagonist (e.g., cesikumab); b) at least one capable of detecting HLA-DRB1*04 duality a probe for the presence of at least one dual gene in the genome; c) instructions for using the probe to analyze the presence of at least one dual gene in the biological sample from the patient; d) with respect to the presence of at least a biological sample from the patient a dual gene is administered to the patient with instructions for the IL-17 antagonist; e) a member selected for administration to the patient's IL-17 binding molecule, as appropriate; and f) at least one selected from the group consisting of therapeutically effective amounts selected from Anti-rheumatic agents of the following groups: immunosuppressants, disease-modifying antirheumatic drugs (DMARDs), pain control drugs, steroids, non-steroidal anti-inflammatory drugs (NSAIDs), cytokine antagonists, bone assimilating agents, bone anti-absorbers, and Its combination.

Disclosed herein are kits for predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist (eg, secukinumab) comprising: a) at least one capable of detecting a consensus epitope a probe for the presence of (SE); and b) instructions for using the probe to analyze the presence or absence of SE in a biological sample from the RA patient, wherein the presence of SE indicates that the patient will be treated with an IL-17 antagonist There is an increased likelihood of a response, and the absence of SE indicates that the patient will have a reduced likelihood of responding to treatment with an IL-17 antagonist.

Disclosed herein are kits for treating a patient with RA comprising: a) a therapeutically effective amount of an IL-17 antagonist (e.g., cesikumab); b) at least one probe capable of detecting the presence of SE ; c) instructions for using the probe to analyze the presence or absence of SE in a biological sample from the patient; d) instructions for administering the IL-17 antagonist to the patient if SE is present in the biological sample from the patient e) a member selected for administration to a patient IL-17 antagonist, as appropriate; and f) optionally a therapeutically effective amount of at least one anti-rheumatic agent selected from the group consisting of immunosuppressive agents, disease modifying agents Antirheumatic drugs (DMARDs), pain control drugs, steroids, nonsteroidal anti-inflammatory drugs (NSAIDs), cytokine antagonists, bone assimilating agents, bone anti-absorbers, and combinations thereof.

Disclosed herein is the use of a kit comprising at least one probe capable of detecting at least one of the dual genes of the HLA-DRB1*04 dual genome, which is used to predict that an IL-17 antagonist will be administered to an IL-17 antagonist (eg, a stopper) Kujinumab) has the potential to respond to treatment.

Disclosed herein is the use of a kit comprising at least one probe capable of detecting SE for predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist (eg, secukimumab).

Disclosed herein is the use of at least one probe capable of detecting the presence of at least one dual gene in a HLA-DRB1*04 dual genome in a biological sample from a patient with RA for predicting that the patient will be against IL-17 An antagonist (eg, cesikumab) treatment is responsive, wherein the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist without the presence of the at least one dual The gene indicates that the patient will have a reduced likelihood of responding to treatment with an IL-17 antagonist.

Disclosed herein is the use of at least one probe capable of detecting the presence of SE in a biological sample from a patient having RA for predicting that the patient will be treated with an IL-17 antagonist (eg, cesikumab) The likelihood of a reaction in which the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with an IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will be antagonistic to the IL-17 The likelihood of treatment responding is reduced.

Disclosed herein are kits comprising: a) a pharmaceutical composition comprising an IL-17 antagonist (eg, cesikumab) for treating rheumatoid arthritis (RA) in a patient; and b) describing how to administer A description of the pharmaceutical composition of the patient, wherein the patient is characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome, or a dual gene in the HLA-DRB1*SE dual genome.

In some embodiments, the probe is capable of detecting the presence of at least one dual gene in the HLA-DRB1*SE dual genome.

In some embodiments, the probe is an oligonucleotide that specifically hybridizes to a nucleic acid region encoding at least one dual gene, an antibody that detects a polypeptide product of at least one dual gene, or an equivalent inheritance encoding at least one dual gene An oligonucleotide that specifically hybridizes to a region of the labeled nucleic acid.

In some embodiments, the presence of SE is detected by analyzing the presence of at least one dual gene in the HLA-DRB1*SE dual genome in the biological sample.

In some embodiments, the presence of two dual genes in the HLA-DRB1*SE dual genome in the biological sample indicates that the patient will further increase the likelihood of responding to treatment with an IL-17 antagonist (eg, cesikumab).

In some embodiments, the presence of two dual genes in the HLA-DRB1*04 dual genome in the biological sample indicates that the patient will further increase the likelihood of responding to treatment with an IL-17 antagonist (eg, cesareimumab).

In some embodiments, the presence or absence of at least one dual gene is detected by analyzing an genomic sequence of at least one dual gene in the biological sample, a product of at least one dual gene, or an equivalent genetic marker of at least one dual gene.

In some embodiments, the biological sample is selected from the group consisting of synovial fluid, blood, serum, plasma, urine, tears, saliva, cerebrospinal fluid, white blood cell samples, and tissue samples.

In some embodiments, the patient is a high risk RA patient.

summary

In preferred embodiments of the disclosed methods, uses, pharmaceutical compositions, kits, assays, and therapeutic regimens, the IL-17 antagonist is selected from the group consisting of: a) an IL-17 binding molecule or IL-17 receptor a body-binding molecule; b) cesikumab; c) an IL-17 antibody that binds to the following epitopes in IL-17: Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129; d) IL-17 antibody binding to IL-17 comprising the following epitopes: Tyr43, Tyr44, Arg46, Ala79, Asp80; e) binding to IL- having two mature IL-17 protein chains 17 homodimeric epitope IL-17 antibody comprising Leu74, Tyr85, His86, Met87, ASn88, Val124, Thr125, Pro126, Ile127, Val128, His129 and in another strand on one strand Containing Tyr43, Tyr44, Arg46, Ala79, Asp80; f) an IL-17 antibody that binds to an epitope having two IL-17 homodimers of the mature IL-17 protein chain, the epitope is in one strand Contains Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val12 8, His129 and contains Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other strand, wherein the IL-17 binding K _D molecule is from about 100 pM to 200 pM, and wherein the IL-17 binding activity of the molecule in vivo half-life Is about 4 weeks; and g) an IL-17 antibody comprising an antibody selected from the group consisting of: i) an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 8 (V) _H); ii) comprising the SEQ ID NO: 10 an immunoglobulin light chain variable domain amino acid sequence (V _L) as shown; iii) comprising the SEQ ID NO: 8 the amino acid sequence shown in FIG. and an immunoglobulin V _H domain comprising the SEQ ID NO: immunoglobulin V _L domain of the amino acid sequence shown in FIG. 10; iv) comprising an immunoglobulin V _H domain as shown in the following variations of high regions: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; v) comprising an immunoglobulin V _L domain as shown in the following variations of high regions: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO :6;vi) an immunoglobulin _VH domain comprising a hypervariable region as shown below: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13; vii) comprising a hypervariable as shown below Immunoglobulin V _H domain of the region: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and comprising an immunoglobulin V _L domain as shown in the following variations of high regions: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and viii) comprising the following hypervariable region as shown in the immunoglobulin V _H domain: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, and comprising an immunoglobulin hypervariable regions V of the _L domain as shown in: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

In the above methods, treatment regimens, uses and pharmaceutical compositions, the preferred embodiments use IL-17 binding molecules, even better embodiments use human antibodies against IL-17, and the preferred embodiment uses Sekukin anti.

All patents, published patent applications, publications, references, and other materials mentioned herein are hereby incorporated by reference. Details of one or more embodiments of the invention are set forth in the accompanying description above. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. Other features, objects, and advantages of the invention will be apparent from the description and appended claims. In the specification and the appended claims, the singular includes the plural unless the context clearly dictates otherwise. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains, unless otherwise defined. All patents and publications cited in this specification are hereby incorporated by reference. The following examples are presented to more fully illustrate the preferred embodiments of the invention. The examples are not to be construed as limiting the scope of the invention as disclosed in the appended claims.

Instance Example 1: Treatment of RA with cesikumab (Studies A2101 and F2201)

Example 1.1-A2101 proof of concept study and results

Clinical Study A2101 is a single-dose and multi-dose type with placebo as a control for the safety and pharmacokinetics of serkumumab in patients with active RA who are taking a stable dose of methotrexate Double-blind proof-of-concept research. 96 patients with rheumatoid arthritis and 8 healthy volunteers were included in the trial. The test was divided into 3 fractions and the dose was gradually increased from 1 x 0.3 mg/kg to 2 x 10 mg/kg i.v.

The primary efficacy analysis was based on 52 patients randomized to receive two infusions of 10 mg/kg cesumuzumab (n=26) or placebo (n=26). Efficacy populations include male and female patients with active RA who are taking MTX. The primary endpoint of the study was the ACR20 response rate at week 6. Secondary endpoints included other time points and DAS28 responses. At the primary endpoint of week 6, the ACR20 response rate for the securkinumab 2 x 10 mg/kg group was 46%, compared to 27% for the placebo group [Δ=19%; P value = 0.13]. The ACR50 response rate and ACR70 response rate of the cesikumab group were 27% [Δ=12%] and 8% [Δ=0%], respectively. DAS28 decreased over time, with a decrease in DAS28 in the 2 x 10 mg/kg group of securkinumab compared to the placebo group. The response to treatment is rapid. The percentage of ACR20 response rate in the cesikumab group was 50% at week 4 and the percentage of ACR20 response in the placebo group was 31%, and at week 16 the ACR20 in group AIN4 secukinumab 57 The percentage of response was 54% and the percentage of ACR20 response in the placebo group was 31%. Although the number in the other dose groups was lower, analysis of patients receiving 2 x 1 mg (n = 6) and patients receiving 3 x 1 mg (n = 6) indicated that these doses were also effective in treating RA.

Example 1.2-F2201 Research Design

Eligible patients (ie, meeting the ACR 1987 revised RA classification criteria for at least 3 months) need to present an active RA as defined below at the time of randomization to ensure the ability to detect response to treatment using ACR criteria: 28 In the joint 6 joint tenderness and 28 joints 6 joint swelling and hsCRP 10 mg/L or ESR 28 mm / 1 hour (mm / h). Eligible candidates take MTX for at least 3 months and currently use a weekly stable dose of MTX when selected ( 7.5 mg / week to 25 mg/week for at least 4 weeks. Adult RA patients taking methamphetamine (n=237) were similarly randomized to receive a monthly subcutaneous injection of securginumab 25 mg, 75 mg, 150 mg, 300 mg or placebo. Patients previously exposed to biologics were included in all cohorts (18% to 22%). The primary endpoint was the proportion of patients who achieved the American College of Rheumatology (ACR) 20 at week 16. At week 20 (8th visit), patients who were randomized to receive placebo at week 0 or who received randomized cetuzumab but did not achieve ACR20 response at week 16 were redistributed Subjectively receiving double-blind treatment until week 48, final efficacy assessment was performed at week 52, and follow-up visits were performed at week 60, starting at week 20: patients who received active treatment as responders continued their dosing Program;

‧All patients in the placebo group were converted to active treatment of 150 mg subcutaneous q4wk (once a month), independent of disease activity;

‧ All patients treated as non-responders treated with 25 mg or 75 mg cesomuzumab q4wk were converted to receive 150 mg subcutaneous q4wk;

The non-responders in the ‧150 mg group were converted to receive the next highest dose of -300 mg subcutaneous q4wk;

All patients in the ‧300 mg group maintained their respective doses to assess whether exposure over 16 weeks would induce clinical response in these patients.

The primary efficacy variable was the clinical response to treatment improvement at week 16 based on ACR20 individual disease activity (for information on ACR scores, see Felson et al, (1995) Arthritis Rheum; 38(6): 727-35). The results were assessed by the proportion of patients who achieved the ACR20 response criteria at week 16.

Other measures include ACR50 (at least 3 of the 5 measures of item B (above) achieve 50% improvement and 50% improvement in swelling and tender joint count), ACR70 (at least 3 of 5 measures of item B (above)) A 70% improvement in measurement and a 70% improvement in swelling and tender joint count) and DAS28 (Disease Activity Score -28) (for information on DAS scores, see Fransen et al., (2003) Ann Rheum Dis; 62 (supplement 1) : 10; Prevoo et al., (1995) Arthritis Rheum; 38(1): 44-48). DAS28 is a recognized measure of RA disease activity. The score is calculated by a complex mathematical formula that includes the number of tender and swollen joints (out of a total of 28 joints), red blood cell sedimentation rate (ESR) or hsCRP, and the overall assessment of the patient's overall health (from excellent to very poor Marked by the 100 mm line). A DAS28 score greater than 5.1 indicates active disease, a score less than 3.2 indicates that the disease is well controlled, and a score less than 2.6 indicates remission.

Example 1.3-F2201 Results

Demographic and baseline characteristics were similar across all groups. At week 16, the ACR20 responders (46.9%, 46.5%, and 53.7%, respectively) in the 75 mg, 150 mg, and 300 mg dose groups of the securkinumab were more than the placebo group (36.0%) and Secu Jinmumab 25 mg group (34%). These results were not statistically significant due to a significant and unexplained increase in ACR20 between week 12 (24%) and week 16 (36%) in the placebo group. A clinically relevant decrease in DAS28-CRP was observed in the securkinumab 75 mg-300 mg treatment group compared to the placebo group. Serum CRP levels were significantly lower at week 16 in the cesugumab 75 mg-300 mg group compared with the placebo group (p=0.0012, 0.0081, and 0.0241). The Acer50 and ACR70 showed a consistently greater improvement in ACR50 and ACR70 in the 16-week dose group compared to the placebo group. HAQ in the 150 mg-300 mg group at week 16 The mean reduction in scores from baseline was approximately 4 times that of the placebo group.

By week 24, the ACR20 response was maintained between the 16th week and the 24th week with the 75 mg to 300 mg group treated with cesumumab and the DAS28 CRP response was further improved. The 75 mg to 300 mg ACR20 responder treatment group showed an early improvement in HAQ score over time until week 24. The ACR50 response initially randomized to patients in the respective dose group improved further from 19% to 24% (75 mg), improved from 21% to 25% (150 mg) and improved from 19% to 24% (300 mg) ), one of these patients had an increased dose at week 20; a similar improvement in ACR70 response was observed in the 75 mg to 150 mg group. It was also noted that the ACR20/50/70 response in patients randomized to placebo increased between weeks 16 and 24.

Example 2: Materials and methods for pharmacogenetic analysis in F2201 and A2101

In the search for a drug response (ie, the response of RA patients to cesomumab), we analyzed 40 SNPs of arthritis risk and multiple SNPs of IL-17A exons (15 known SNPs and 5 Unreported SNPs, and patients who genotyped HLA-DRB1 dual genes at weeks 12 and 16 in efficacy data. We additionally used data over the 16th week to verify the correlation between the specific HLA-DRB1 dual gene and the cesomumab response. Our results indicate that the high cesumab reaction population is present in individuals with a specific HLA-DRB1 dual gene. The following discussion is about our analysis of the HLA-DRB1 dual gene.

Example 2.1: Samples and Processing

The DNA of 149 informed consent patients participating in the F2201 study was genotyped. 121 patients receiving securkinumab were used in pharmacogenetic (PG) analysis. Genotyping the DNA of 32 informed consent patients enrolled in the A2101 study, including 20 patients receiving 2 x securkinab 3.0 mg/kg or 2 x securkinab 10.0 mg/kg Patients and 12 patients who received a matching placebo (FIR efficacy endpoints were only measured in these groups). Thirty Caucasian patients were used in the genetic analysis, including 18 patients receiving securkinumab and 12 patients receiving placebo.

Blood samples from patients were collected at individual test sites and then shipped to Covance (Indianapolis, USA and Geneva, Switzerland). Genomic DNA from each patient was extracted from the blood by Covance using the PUREGENE D-50K DNA isolation kit (Gentra, Minneapolis, MN, USA) and shipped to Novartis for genotyping. Test HLA-DRB1*SE dual genome (HLA-DRB1*01:01, HLA-DRB1*01:02, HLA-DRB1*04:01, HLA-DRB1*04:04, HLA-DRB1*04:05, HLA -DRB1*04:08, HLA-DRB1*10:01, HLA-DRB1*14:02) and HLA-DRB1*04 dual genome (four-digit dual gene) are associated with different responses to cesomumab treatment Sex. The HLA-DRB1*04 dual genome was also genotyped in A2101 to replicate the correlation with the crustum mAb observed in the F2201 sample.

All samples of the F2201 study (n=150) were subjected to sequence-specific oligonucleotide hybridization (SSO), which typically produced low-resolution results for HLA-pair genes (2-digit dual gene names). In short, by using LABType SSO experiments were performed on HD Class II DRB1 typing tests (One lambda, Inc, CA) and Luminex IS200 instruments. By using HLA Fusion 2.0 software (One Lambda) analysis data. Of the first and second batches, one failed to produce results. For the remaining 149 samples, if it is clear at this experimental stage or the ambiguity can be resolved by excluding rare dual genes (see rare_alleles_2_28_0, available at bioinformatics.nmdp.org/HLA/Biannual_Rare_Allele_List/index.html) , then specify the final high-resolution result (4 digits of the dual gene name). Sequence-based typing (SBT) and sequence-specific primer (SSP) PCR were further performed on samples whose genotypes were not resolved at the 4-digit level. Briefly, SBT experiments were performed by using the HLA-DRB1 SBT package (Abbott Molecular, IL) and the SBTexcellerator HLA DRB1 kit (Qiagen, Netherlands). The sequencing products were separated on a 3730xl Genetic Analyzer (Applied Biosystems, CA) and the fluorescence of the dye was analyzed to determine the DNA sequence. The genotype was analyzed and assigned by using SBTengine software 2.12.1.0 (Genome Diagnostics, Netherlands). SSP was performed using the Olerup SSP kit (Olerup, PA). The resulting PCR amplicons were analyzed by separation on an agarose gel in terms of its length compared to size standards. The final dual genotype was specified by comparison to the SSP worksheet provided by Olerup.

Of the samples of the A2101 study (n=32), only HLA-DRB1*04 was further resolved and reported at high resolution using methods as described above, while other dual genotypes were reported as SSO data output.

Example 2.2: Statistical Analysis

In general, the statistical model of pharmacogenetic analysis is based on the model used in clinical trial analysis, adding the genotypes of the variables (eg, HLA dual genes) and other covariates (if applicable). All variables are tested individually, ie only one variable is included at a time in the model. All HLA dual genes are tested for clinical endpoints using a standard additive effect code: the individual is coded as 0, 1 or 2 for the HLA dual gene, depending on the number of HLA dual gene copies carried by the individual. All correlation tests for the additive dual gene effect are two-tailed single-point tests.

Race is a common distracting factor in genetic correlation studies. Approximately 87% of the 121 patients treated with securkinumab in F2201 were Caucasian. Two models were tested according to race in the F2201 sample: a) included all patients treated with cesumumab (N=121) and adjusted for race; b) included only Caucasian patients treated with secukinumab And not adjusted for race (N = 105). Since most A2101 patients (about 94%) are Caucasian, only 30 Caucasian patients are eligible for analysis in A2101.

Genetic analysis was performed using only F2201 samples from patients treated with cesumumab (N=121). The null hypothesis assumes that the coefficient of the genotype variable is equal to zero and presents the corresponding p-value. Rejecting the null hypothesis would mean the conclusion that the genotype is a predictor of response to securizumab as measured by a specific clinical endpoint.

Genetic analysis (N=30) was performed using A2101 samples from Caucasian patients receiving securginumab and placebo. Other items are included in the genetic analysis model in A2101: the interaction between this indicator variable and genotype variables in the treatment group. The null hypothesis is that the coefficient of interaction between the therapeutic index and the genotype variable is equal to zero and presents a corresponding p-value. Rejecting the null hypothesis would mean the conclusion that the genotype is a predictor of the extent to which a patient's response to securizumab is better than the placebo response as measured by a specific clinical endpoint.

Example 2.2.1: Exploratory PG analysis in F2201

Example 2.2.1.1: DRB14 digital dual gene in all F2201 patients treated with cesumumab

The genotype of the HLA DRB1 gene in 149 patients in the F2201 study as described above was obtained at 4 digit resolution. The correlation between all DRB1 4 digit dual genes and the cesomumab reaction was tested.

All statistical tests were performed with SAS. The ANCOVA model (SAS 9.2 PROC GLM) was used to analyze the efficacy variable DAS28 obtained by CRP at week 12/week 16 and the logistic regression model (SAS 9.2 PROC LOGISTIC) was used to analyze week 12/16 The weekly variable ACR20, which has an efficacy endpoint as a dependent variable, is a DRB1 4 digit dual gene (as encoded above) as an independent variable (fixed effect) and has the following fixed effect covariates:

‧gender

‧Drug group or drug concentration, two models for each test

‧Baseline weight (included only when adjusting the dosing group for baseline weight)

‧ Race (included only when using all patients treated with cesomumab for analysis)

‧ Baseline DAS28 obtained by CRP

‧Baseline RF (known to be associated with efficacy response)

‧ Baseline anti-CCP antibody (known to be associated with efficacy response)

‧ Baseline CRP (known to be associated with efficacy response)

‧Baseline ESR (known to be associated with efficacy response)

The last observation was not carried over to estimate the missing value. Exclude the polymorphism of the frequency <5% in the sample.

Example 2.2.1.2: HLA-DRB1*04 dual gene in all F2201 patients treated with cesumumab

A similar test was performed for the cesikumab reaction at week 12/week 16 as described above, with the exception of the two-tailed single-point test for the additive dual gene effect test HLA-DRB1*04 dual genome and Sekukin Resistance to reaction.

The correlation between the HLA-DRB1*04 dual genome and the observed cesomumab response in the data over the 16th week was also analyzed to verify the results at week 12/week 16. A three-part pharmacogenetic analysis of data over the 16th week was performed.

The first part of the analysis was to test the correlation between DRB1*04 and the efficacy at the 16th week in patients randomized to the cesikumab group at baseline (N=117). At the 20th week, ACR20 non-responders were converted to receive higher doses of securinumab, which may cause a loss of assay power for this analysis. Part II analysis was performed on patients treated with cesumimumab (N=54) defined as ACR20 responders at week 16. This analysis should not be affected by potential interference due to changes in the dosage regimen as this subgroup maintains the original administration. Part III analysis was performed on the placebo group. The placebo group was switched to receive securginum 150 mg at week 20. Therefore, week 20 was used as the baseline in this analysis.

Example 2.2.1.3: HLA-DRB1*04 dual gene in patients with F2201 who were not treated with anti-TNF antibody and who were treated with securginumab

Similar tests were performed as described above with the exception of patients who were previously treated with anti-TNF antibodies.

Example 2.2.1.4: HLA-DRB1*SE dual gene in all F2201 patients treated with cesumumab

A similar test was performed as described above, except that the DRB1 SE dual genome was tested for the additive dual gene effect using a two-tailed single-point test (HLA-DRB1*01:01, HLA-DRB1*01:02, HLA-DRB1*04:01 , HLA-DRB1*04:04, HLA-DRB1*04:05, HLA-DRB1*04:08, HLA-DRB1*10:01, HLA-DRB1*14:02) Correlation with Securkinumab Sex.

Example 2.2.2: Testing the correlation between HLA-DRB1*04 and the cesikumab reaction in A2101

Example 2.2.2.1: Main purpose - Test the effect of HLA-DRB1*04 on DAS28 (interaction term) at day 43 (week 6)

The genotype of the HLA DRB1 gene of 32 informed consent patients in the A2101 study as described above was obtained at 2 digit resolution. By comparing a patient carrying at least one HLA-DRB1*04 dual gene (coded as 1) with a patient not carrying the dual gene (encoded as 0) and determining the cesikumab reaction between the two groups Whether it is significantly different to test the correlation of the dual genome HLA-DRB1*04. A dominant genetic model was used in this test because only one patient in this small sample carried two HLA-DRB1*04 dual genes. Genetic analysis was performed using A2101 samples from patients treated with cesumimumab and patients receiving placebo.

Statistical testing was performed with SAS. The efficacy variable used in this test is the change in DAS28 from baseline on day 43 (assessment of blind observers). Efficacy variables were analyzed using the ANCOVA model (SAS 9.2 PROC GLM), where the efficacy endpoint was used as the dependent variable, the genotype variable of the HLA-DRB1*04 carrying state (as encoded above) as the independent variable (fixed effect), the treatment group (whether the patient is Randomized to receive an indicator of cesomumab or placebo) and baseline DAS28 as a fixed-effect covariate, and the following additional items were included in the model: the interaction between the indicated variables and genotype variables in this treatment group .

Example 2.2.2.2: Secondary purpose

Test HLA-DRB1*04 for ACR20/ on Day 43 (Week 6) ACR50 (interaction term) effect

A similar test was performed as described above with the exception that ACR20/ACR50 was used as the efficacy variable at day 43 (week 6). Two functional variables were analyzed separately using a logistic regression model (SAS 9.2 PROC LOGISTIC).

Test the effect of HLA-DRB1*04 on DAS28/ACR20/ACR50 (interaction term) during repeated visits (week 3, week 4, week 5, week 6)

The generalized estimation equation (GEE) method (SAS 9.2 PROC GENMOD) was used to analyze the efficacy endpoint of HLA-DRB1*04 for repeated visits (week 3, week 4, week 5, week 6). The effect of ACR20/ACR50 (interaction term) to explain the correlation between the repeated measurements. The model has a efficacy endpoint as a dependent variable, the HLA-DRB1*04 dual gene as an independent variable (fixed effect), the treatment group (whether the patient was randomized to receive an indicator of securginum or placebo) and baseline DAS28 as a fixation The effect covariates, and the following other items are included in the model: this treatment group indicates the interaction between the variables and the genotype variables. The evaluation structure for the correlation between the results at different times is autoregressive (TYPE=AR in SAS 9.2 PROC GENMOD).

Example 3: Results of pharmacogenetic analysis in F2201 (week 12 and week 16)

Example 3.1: Exploratory PG Analysis in F2201

Example 3.1.1: Testing the effect of HLA-DRB1 dual gene on the reaction of sequocumab

Example 3.1.1.1: Effect of HLA-DRB1 dual gene on the response to all cetuzumab in patients treated with cetuzumab

Among the 4-digit DRB1 dual genes, the p-value of HLA-DRB1*0401 was the best, and the additive model was used to adjust the following for the correlation with the ACR20 of all 121 patients at week 12. = 0.0394: drug concentration, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

Example 3.1.1.2: Effect of HLA-DRB1*04 dual gene on the response of all F2201 patients treated with secukimumab to securizumab

Of the 121 patients treated with cesumumab in F2201, 48% had at least one HLA-DRB1*04 dual gene. As shown in Table 5 , a higher percentage of patients with at least one HLA-DRB1*04 dual gene achieved ACR20 and ACR50 at weeks 12 and 16 after treatment with serkuumumab.

Get the following results:

‧ Using the additive model, adjusted for the following, the p-value obtained for the correlation with ACR20 of all 121 patients at week 12 = 0.0032, odds ratio (OR) = 2.78: drug concentration, gender, ethnicity Baseline DAS28, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR obtained from CRP.

‧ Use the addition model to adjust for the correlation with DAS28 for all 121 patients at week 16 with the addition of the following values = 0.0046, β = -0.49: drug concentration, gender, ethnicity, baseline obtained from CRP DAS28, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

‧ Using the additive model, the p-value obtained for the correlation with DAS28 of all 121 patients at week 12 was adjusted for the following = 0.0082, β = -0.43: drug concentration, gender, ethnicity, baseline obtained from CRP DAS28, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

Table 5 shows the percentage of patients treated with cesumumab at the given endpoint (ACR20, ACR50, ACR70). HLA-DRB1*04-=Phase without HLA-DRB1*04 dual gene; HLA-DRB1*04+ = patient with at least one HLA-DRB1*04 dual gene.

The effect of the HLA-DRB1*04 dual gene pair on response to treatment with secukinumab can also be seen in Figure 1 . At week 12, the same type of contiguous individuals (individuals carrying two HLA-DRB1*04 dual gene copies) had the highest ACR20 response rate, followed by heterozygous individuals (an individual carrying a HLA-DRB1*04 dual gene copy). Similar results are seen in Figure 2 , which shows the cumulative effect of the HLA-DRB1*04 dual gene on the percentage of patients who achieved ACR50 with cesibumab at Weeks 12 and 16. At week 12 and week 16, the same type of HLA-DRB1*04 individuals had the highest ACR50 response rate, followed by heterozygous individuals. A similar effect was observed for the DAS28 score at Week 12 and Week 16 ( Figure 3 ), which demonstrates the additive effect of the HLA-DRB1*04 dual gene on the DAS28 score of patients treated with secukinumab. At week 12 and week 16, HLA-DRB1*04 homozygous individuals had the lowest DAS28 score, followed by heterozygous individuals.

As shown in Table 6 , the HLA-DRB1*04 dual gene was significantly lower in DAS28 (at increased response) and placebo (MTX) in patients treated with all doses of secerumab at weeks 12 and 16. The response is reduced. In addition, the HLA-DRB1*04 dual gene was associated with a percentage increase in ACR20 and a decrease in response to placebo (MTX) in patients treated with all doses of serecumab at week 12.

Table 6 shows changes in DAS28_CRP scores at week 12 and week 16 for all patients due to carrier/non-carrier status and treatment (placebo, 25 mg, 75 mg, 150 mg, or 300 mg secerumab) And the percentage of patients who achieved ACR20 or ACR50.

Example 3.1.1.3: HLA-DRB1*04 dual gene pair previously not Effect of anti-TNF antibody on F2201 in patients treated with securkinumab

A total of 15% of eligible F2201 patients were previously treated with anti-TNF therapy. To test whether patients who were refractory to TNF therapy (refractory) were more likely to be cured with serkumuzumab, an analysis similar to that described above was performed by excluding patients previously treated with anti-TNF therapy. As shown in Table 7 , the correlation between HLA-DRB1*04 and cesikumab response observed in the subgroup not treated with anti-TNF antibody was similar to that observed for all patients.

Table 7 shows patients who were not previously treated with cesumumab at the 12th week for carriers/non-carriers and treatment (placebo, 25 mg, 75 mg, 150 mg, or 300 mg cesikumab) Changes in DAS28_CRP score at week 16 and percentage of patients achieving ACR20 or ACR50.

Example 3.1.1.4: HLA-DRB1*SE dual gene pair F2201 Effect of cesikumab reaction in patients treated with kujiumab

As shown in Table 8 , a higher percentage of patients with at least one HLA-DRB1*SE dual gene achieved ACR20, ACR50, and ACR70 at weeks 12 and 16 after treatment with serkuumumab.

Get the following results:

‧ Use the addition model to adjust the following for the correlation with the ACR50 of all 121 patients at week 12: p value = 0.010, OR = 3.92: drug concentration, gender, ethnicity, baseline DAS28 obtained from CRP Baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

‧ Using the dominant model, adjusted for the following, for the correlation with the ACR50 of all 121 patients at week 12, the p-value = 0.027, OR = 13.31: drug concentration, gender, ethnicity, baseline obtained from CRP DAS28, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

‧ Using the dominant model, adjusted for the following, the p-value obtained for the correlation with DAS28 of all 121 patients at week 16 = 0.025, β = -0.57: drug concentration, gender, ethnicity, obtained by CRP Baseline DAS28, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

Table 8 shows the percentage of patients treated with cesumumab at the given endpoint (ACR20, ACR50, ACR70). DRB1*SE-=Phase without HLA-DRB1*SE dual gene; HLA-DRB1*SE+ = patient with at least one HLA-DRB1*SE dual gene.

As shown in Table 9 , the HLA-DRB1*SE dual gene and the patients treated with cesomumab in the 150 mg and 300 mg dose groups at weeks 12 and 16 were significantly lower in DAS28 and in the placebo group ( The response of MTX) is reduced. HLA-DRB1*SE dual gene and percentage of ACR20 increased in patients treated with cesumumab in the 75 mg to 300 mg group at week 12 and week 16 and decreased response to placebo (MTX) Related. The HLA-DRB1*SE dual gene was also associated with an increase in the percentage of ACR50 achieved by patients treated with secerumab in the 75 mg to 300 mg group at weeks 12 and 16.

Table 9 shows all patients with HLA-DRB1*SE dual gene carrier/non-carrier status and treatment (placebo, 25 mg, 75 mg, 150 mg, or 300 mg cesikumab) at week 12 and Changes in DAS28_CRP score at 16 weeks and percentage of patients achieving ACR20 or ACR50.

Example 4: HLA-DRB1*04 dual gene analysis in the A2101 study (week 7)

Example 4.1: Main purpose results

HLA-DRB1*04 dual gene carrier/non-carrier status was significantly associated with cesikumab response using a blinded observer assessment of DAS28 on day 43 as a measure of efficacy, with The action term HLA-DRB1*04×treatment p=0.042. As shown in Table 10 , the HLA-DRB1*04 carrier/non-carrier status was also identified as the response to sexeuzumab using the blind observer assessment of DAS28 on day 43 as a measure of efficacy. A subgroup of reactions to placebo (MTX). In addition, the response of the HLA-DRB1*04 dual gene to the cesikumab group was increased and the response in the placebo group (MTX) was reduced using the DAS28 investigator's rating as the efficacy measure on day 43. Small correlation.

Table 10 shows changes in DAS28_CRP scores at day 43 for all Caucasian patients due to HLA-DRB1*04 dual gene carrier/non-carrier status and treatment (placebo or serkuumumab) and achieving ACR20 or ACR50 Percentage of patients.

Example 4.2: Secondary purpose results

As shown in Table 11 , the percentage of ACR20/ACR50 in the HLA-DRB1*04+ subgroup was higher in patients treated with cesumumab. For Day 43 ACR20/ACR50, the genotype* treatment interaction term was not statistically significant and was most likely due to the lack of assay power at the endpoint of the binary efficacy. However, when the GEE method was used to interpret the correlation between the repetitive measurements (week 3, week 4, week 5, week 6), a significant genotype* therapeutic interaction was observed for ACR20, where p=0.025 . The effect of HLA-DRB1*04 on the DAS28, ACR20 and ACR50 responses of the A2101 sample persisted over time.

Table 11 shows the percentage of patients treated with secerumab treated to a given endpoint (ACR20, ACR50, ACR70). HLA-DRB1*04-=Phase without HLA-DRB1*04 dual gene; HLA-DRB1*04+ = patient with at least one HLA-DRB1*04 dual gene.

Example 5: Results of pharmacogenetic analysis in F2201 over week 16

Example 5.1: Randomization to all of the cesikumab groups at baseline Patient analysis

We tested the association between HLA-DRB1*04 and the efficacy of patients randomized to the cesikumab group at baseline (N=117) over the 16th week. HLA-DRB1*04 was observed to be associated with a better cesbaunumab response over time in terms of ACR50 ( Figure 4 ), ACR20 ( Figure 5 ), and DAS28 score ( Figure 6 ).

Get the following results:

‧ P = 0.0057, odds ratio (OR) obtained for the association of ACR50 at week 52 with 117 patients randomized to the cesumumab group at baseline using the additive model = 3.97: dosing group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

‧ Using the additive model, adjusted for the following, the p-value obtained for the association of DAS28 at week 52 with 117 patients randomized to the cesumuzumab group at baseline = 0.0023, β =- 0.60: dosing group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

As shown in Table 12 , HLA-DRB1*04 dual gene was associated with a greater decrease in DAS28 (indicating an increased response) at week 52 in patients treated with all doses of serecumab except for the 25 mg dose. In addition, the HLA-DRB1*04 dual gene was associated with an increase in the percentage of ACR50 achieved at week 52 by patients treated with all doses of serecumab. Similar findings were observed in a subgroup analysis of patients who were not treated with anti-TNF antibodies. Data beyond the 16th week supported the correlation between DRB1*04 and response observed in the 12th and 16th week data. Please note that at the 20th week, ACR20 non-responders were converted to receive higher doses of Securkinumab, which may cause a loss of assay power for this analysis.

Table 12 : Study of Week 52 results for all cesumuzumab patients in F2201.

Example 5.2: Subgroup analysis of patients treated with cesumumab at the 16th week defined as ACR20 responders

This subgroup analysis was performed on patients treated with cesumumab at the 16th week defined as ACR20 responders. The goal of this analysis was to test the correlation between DRB1*04 and patients treated with cesikumab (N=54) who were defined as ACR20 responders at week 16 over the 16th week. This analysis should not be affected by potential interference due to changes in the dosage regimen as this subgroup maintains the original administration. HLA-DRB1*04 was observed to be associated with a better cesbaunumab response over time in terms of ACR50 ( Figure 7 ), ACR20 ( Figure 8 ), and DAS28 score ( Figure 9 ).

Get the following results:

‧ Using the additive model, adjusted for the following, p=0.0082 for the correlation of ACR50 at week 52 with 54 patients defined as ACR20 at week 16, the odds ratio (OR) = 70.81 : Administration group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

‧ Using the additive model, adjusted for the following, the p-value obtained for the DAS28 at week 52 for 54 patients who were defined as ACR20 responders at week 16 = 0.013, β = -0.81: Drug group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

As shown in Table 13 , patients with HLA-DRB1*04 dual gene and all doses of cesumumab treated with a dose other than 25 mg defined as ACR20 responders at week 16 were at week 52 at DAS28. More reduction (indicating an increase in response) is associated. In addition, the HLA-DRB1*04 dual gene was associated with an increase in the percentage of ACR50 achieved at week 52 by patients treated with all doses of serecumab. Similar findings were observed in a subgroup analysis of patients who were not treated with anti-TNF antibodies. This genetic analysis supports the correlation between DRB1*04 and response observed in the 12th and 16th week data.

Table 13 : Week 52 results for subgroup analysis of patients treated with cesumimumab as ACR20 responders at week 16

Example 5.3: Subgroup analysis of placebo group (transformed to receive 150 mg cesikumab at week 20)

This subgroup analysis was performed on the placebo group. The placebo group was switched to receive securginum 150 mg at week 20. Information for the 20th week is not available. Thus, week 16 was used as the baseline in this analysis. The goal of this analysis was to reproduce the correlation between DRB1*04 and response observed in the cesikumab group in an independent sample.

Get the following results:

In all placebo groups (N=25, Table 14 ):

‧ Use the addition model to adjust the following for the correlation with the ACR50 at week 52 for all placebo groups (N=25), p=0.30, odds ratio (OR)=11.59: race and by CRP Obtain the baseline DAS28 (omit other covariates to make the model likely to converge).

‧ Using the additive model, adjusted for the following, the p-value obtained for the DAS28 at week 52 for 54 patients who were defined as ACR20 responders at week 16 = 0.92, β = -0.053: Drug group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

In the placebo group previously not treated with anti-TNF antibody (N=20, Table 14 ):

‧ Only 2 patients in this subgroup achieved ACR50 and therefore cannot calculate p-values for correlation with ACR50.

‧ Using the additive model, adjusted for the following, the p-value obtained for the DAS28 at week 52 for 54 patients who were defined as ACR20 responders at week 16 = 0.47, β = -0.53: Drug group, gender, ethnicity, baseline DAS28 obtained from CRP, baseline RF, baseline anti-CCP antibody, baseline CRP, and baseline ESR.

In all placebo groups (N=25), there was no consistent evidence of a correlation with the response to cesomumab in ACR50 ( Figure 10 ) or ACR20 ( Figure 11 ). Although the DAS28 score for HLA-DRB1*04 carriers was higher from the 16th week to the 52nd week ( Fig. 12 ), this correlation trend appears to be mainly caused by baseline interference factors. It is unclear why the correlation between HLA-DRB1*04 and cesikumab response was not replicated in all placebo subgroup analyses. However, for recurring purposes, the sample size is extremely small. In addition, at week 20, the placebo group was switched from placebo (MTX) to receiving securginumab 150 mg. These factors, together with the unexpectedly high placebo effect observed at week 16 in clinical studies, may bias the subgroup analysis of Example 5.3.

Interestingly, the trend in the subset of patients without anti-TNF antibody (N=20) was generally consistent with the cesikumab group (p-value was not significant). It was observed that HLA-DRB1*04 was associated with a better cesbaunumab response over time in patients treated with no anti-TNF antibody ( Figures 13-15 ). Although the placebo group did not demonstrate a correlation between HLA-DRB1*04 and cesumumab response, a subgroup of this subgroup (ie, patients without anti-TNF antibody therapy) showed a trend of relevance. It may mean that there is a specific value when using cesikumab in patients who are not treated with anti-TNF antibodies. However, the small sample size used in Example 5.3 should be noted when interpreting any of these groups (N=25 for all placebo groups; N=20 for placebo groups without anti-TNF antibody treatment).

Table 14: Week 52 results for a subgroup analysis of the placebo group (converted to receive 150 mg secerumab at week 20).

In summary, the information provided herein supports the correlation between HLA-DRB1*04 and/or HLA-DRB1*SE status and response to serecumab in RA patients as measured by DAS28 and ACR scores. This study is currently validated in a prospective clinical trial. It is worth noting that SE has previously been shown to not predict response to biological agents, especially anti-TNF factor treatments (such as etanercept to infliximab) (Potter et al., (2009) Ann. Rheum. Dis. 68:69 -74). Potter et al. showed no correlation between the anti-TNF response and the risk-bearing gene carrying either of the two recognized RA-sensitive factors (SE or PTPN22 ). Thus, we have surprisingly determined that there is a significant correlation between the HLA-DRB1*04 and/or HLA-DRB1*SE status and the response of RA patients to securizumab.

Figure 1 shows the effect of the HLA-DRB1*04 dual gene pair on the ACR20 response at week 12 for all patients randomized to the cesumuzumab group at baseline.

Figure 2 shows the effect of HLA-DRB1*04 dual gene pair ACR50 at week 12 and week 16 on all patients randomized to the cesumuzumab group at baseline.

Figure 3 shows the effect of the HLA-DRB1*04 dual gene pair on the DAS28 score at week 12 and week 16 for all patients randomized to the cesikumab group at baseline.

Figure 4 shows the effect of HLA-DRB1*04 dual gene pair ACR50 from week 12 to week 52 on all patients randomized to the cesikumab group at baseline.

Figure 5 shows the effect of HLA-DRB1*04 dual gene pair ACR20 from week 12 to week 52 on all patients randomized to the cesicumab group at baseline.

Figure 6 shows the effect of HLA-DRB1*04 dual gene pair DAS28 scores from week 12 to week 52 on all patients randomized to the cesikumab group at baseline.

Figure 7 shows the effect of HLA-DRB1*04 dual gene on ACR50 from week 12 to week 52 of patients treated with cesumumab at the 16th week defined as responder (ACR20).

Figure 8 shows the effect of HLA-DRB1*04 dual gene on ACR20 from week 12 to week 52 of patients treated with cesumumab at the 16th week defined as responder (ACR20).

Figure 9 shows the effect of the HLA-DRB1*04 dual gene pair on the DAS28 score from week 12 to week 52 for patients treated with cesicumab at the 16th week as responder (ACR20).

Figure 10 shows the effect of HLA-DRB1*04 dual gene pair ACR50 from week 12 to week 52 on all patients randomized to placebo at baseline.

Figure 11 shows the effect of HLA-DRB1*04 dual gene pair ACR20 from week 12 to week 52 on all patients randomized to placebo at baseline.

Figure 12 shows the effect of HLA-DRB1*04 dual gene pair DAS28 scores from week 12 to week 52 on all patients randomized to placebo at baseline.

Figure 13 shows the effect of HLA-DRB1*04 dual gene pair ACR50 from week 16 to week 52 on patients who were previously randomized to placebo in the placebo group who were not treated with anti-TNF antibody.

Figure 14 shows the effect of HLA-DRB1*04 dual gene pair ACR20 from baseline to week 52 on patients who were previously randomized to placebo in the placebo group treated with no anti-TNF antibody.

Figure 15 shows the effect of HLA-DRB1*04 dual gene pair DAS28 scores from baseline to week 52 on patients who were previously randomized to placebo in the placebo group who were not treated with anti-TNF antibody.

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Claims (40)

A kit for predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist comprising a) at least one capable of detecting at least one dual gene in the HLA-DRB1*04 dual genome a probe present; and b) instructions for using the probe to analyze the presence of the at least one dual gene in a biological sample from the RA patient, wherein the presence of the at least one dual gene indicates that the patient will antagonize the IL-17 There is an increased likelihood that the treatment will respond, and the absence of the at least one dual gene indicates that the patient will have a reduced likelihood of responding to treatment with the IL-17 antagonist. A kit for treating a patient having RA comprising a) a therapeutically effective amount of an IL-17 antagonist; b) at least one capable of detecting the presence of at least one dual gene in the HLA-DRB1*04 dual genome a probe; c) instructions for using the probe to analyze the presence of the at least one dual gene in a biological sample from the patient, d) administering the at least one dual gene in the biological sample from the patient a patient's instructions for the IL-17 antagonist; e) a member selected for administration to the IL-17 antagonist of the patient, as appropriate; and f) optionally at least one selected from the group consisting of Anti-rheumatic agents: immunosuppressants, disease-modifying anti-rheumatic drugs (DMARDs), pain control drugs, steroids, non-steroidal anti-inflammatory drugs (NSAIDs), cytokine antagonists, bone assimilator, bone Anti-absorbents and combinations thereof. A kit for predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist comprising a) at least one probe capable of detecting the presence of SE; and b) using the probe The needle analyzes instructions for the presence or absence of the SE in the biological sample of the RA patient, wherein the presence of the SE indicates that the patient will have an increased likelihood of responding to treatment with the IL-17 antagonist without the presence of the SE indication This patient will have a reduced likelihood of responding to treatment with this IL-17 antagonist. A kit for treating a patient with RA comprising a) a therapeutically effective amount of an IL-17 antagonist; b) at least one probe capable of detecting the presence of SE; c) from analyzing using the probe a specification for the presence or absence of the SE in the biological sample of the patient, d) instructions for administering the IL-17 antagonist to the patient if the SE is present in the biological sample from the patient; e) optionally a component for administering the IL-17 antagonist to the patient; and f) optionally a therapeutically effective amount of at least one anti-rheumatic agent selected from the group consisting of an immunosuppressive agent, a disease-modifying antirheumatic drug ( DMARD), pain control drugs, steroids, non-steroidal anti-inflammatory drugs (NSAIDs), cytokine antagonists, bone assimilating agents, bone anti-absorbers, and combinations thereof. The kit of any one of claims 3 to 4, wherein the probe is capable of detecting the presence of at least one dual gene in the HLA-DRB1*SE dual genome. The kit of any one of claims 1 to 5, wherein the probe is an oligonucleotide that specifically hybridizes to a nucleic acid region encoding the at least one dual gene, and the polypeptide product of the at least one dual gene is detectable An antibody, or an oligonucleotide that specifically hybridizes to a nucleic acid region encoding an equivalent genetic marker of the at least one dual gene. An IL-17 antagonist for the treatment of RA, characterized by: a) obtaining a biological sample from a RA patient; b) analyzing the presence or absence of at least one dual gene in the HLA-DRB1*04 dual genome in the biological sample And c) administering the therapeutically effective amount of an IL-17 antagonist to the RA patient if the at least one dual gene is present in the biological sample. An IL-17 antagonist for the treatment of RA, characterized by: a) analyzing the presence or absence of at least one dual gene in the HLA-DRB1*04 dual genome in a biological sample from a RA patient; and b) if The presence of the at least one dual gene in the biological sample is administered to the RA patient in a therapeutically effective amount of an IL-17 antagonist. An IL-17 antagonist for the treatment of RA, characterized by: a) obtaining a biological sample from a RA patient; b) analyzing the presence or absence of SE in the biological sample; and c) if the biological sample is present at least A dual gene is administered to a therapeutically effective amount of an IL-17 antagonist in the RA patient. An IL-17 antagonist for the treatment of RA, characterized by: a) analyzing the presence or absence of SE in a biological sample from a RA patient; and b) if the at least one dual gene is present in the biological sample, A therapeutically effective amount of an IL-17 antagonist is administered to the RA patient. The use of any one of claims 9 to 10, wherein the presence or absence of the SE is detected by analyzing the presence or absence of at least one of the dual genes in the HLA-DRB1*SE dual genome in the biological sample. The use of any one of claims 7 to 8 or 11, wherein the genomic sequence of the at least one dual gene, the product of the at least one dual gene, or the equivalent genetic marker of the at least one dual gene in the biological sample is analyzed To detect the presence or absence of the at least one dual gene. An IL-17 antagonist for the treatment of RA, characterized by: a) selecting a patient with RA for treatment based on the RA patient having at least one dual gene in the HLA-DRB1*04 dual genome; and b) administering The RA patient has a therapeutically effective amount of an IL-17 antagonist. An IL-17 antagonist for treating a patient having RA, characterized by administering the IL-17 antagonist to a patient having at least one of the dual genes in the HLA-DRB1*04 dual genome. An IL-17 antagonist for treating a patient having RA, characterized in that the IL-17 antagonist is administered to a patient selected for treatment based on at least one dual gene having a HLA-DRB1*04 dual genome . An IL-17 antagonist for the treatment of RA, characterized by: a) selecting a patient with RA for treatment based on the patient having RA; and b) administering a therapeutically effective amount of an IL-17 antagonist to the patient with RA . An IL-17 antagonist for treating a patient having RA, characterized in that the IL-17 antagonist is to be administered to a patient having SE. An IL-17 antagonist for treating a patient having RA, characterized in that the IL-17 antagonist is administered to a patient selected for treatment based on having SE. The use according to any one of claims 13 to 18, wherein the patient is in need of intravenous administration of three doses of about 10 mg/kg of the IL-17 antagonist, each of which is delivered every other week. The use of any one of claims 13 to 18, wherein the patient is administered subcutaneously from about 75 mg to about 300 mg twice a month, once a month, once every two months, or once every three months. The IL-17 antagonist. Use of an IL-17 antagonist for the manufacture of a medicament for treating a patient having RA, wherein the patient has at least one dual gene in the HLA-DRB1*04 dual genome. Use of an IL-17 antagonist for the manufacture of a medicament for treating a patient having RA, wherein the patient is selected for treatment based on having at least one dual gene in the HLA-DRB1*04 dual genome. Use of an IL-17 antagonist for the manufacture of a medicament for treating a patient having RA, wherein the RA patient has SE. Use of an IL-17 antagonist for the manufacture of a medicament for treating a patient having RA, wherein the RA patient is selected for treatment based on having SE. A method of determining the responsiveness of a patient having RA to treatment with an IL-17 antagonist comprising: a) analyzing a biological sample from the patient to determine the presence or absence of at least one of the HLA-DRB1*04 dual genomes a dual gene; and b) if the presence of the at least one dual gene is detected in the sample, the patient is designated to be responsive to treatment with the IL-17 antagonist. A method of determining the responsiveness of a patient having RA to treatment with an IL-17 antagonist comprising: a) analyzing a biological sample from the patient to determine the presence or absence of SE; and b) if in the sample Upon detection of the presence of the at least one dual gene, the patient is designated to respond to treatment with the IL-17 antagonist. A method of generating a form of transmitable information for predicting responsiveness to treatment with an IL-17 antagonist in a patient having RA comprising: a) analyzing a HLA-DRB1*04 dual genome in a biological sample from the patient The presence or absence of at least one dual gene; and b) materializing the results of the analysis step into a form of transportable information. A method of generating a form of transmittable information for predicting responsiveness to treatment with an IL-17 antagonist in a patient having RA comprising: a) analyzing the presence or absence of SE in a biological sample from the patient; b) The result of the analysis step is embodied in the form of transportable information. A method of predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist comprising analyzing at least one duality of the HLA-DRB1*04 dual genome in a biological sample from the patient using an automated analyzer The presence or absence of a gene, wherein the presence of the at least one dual gene indicates that the patient will have an increased likelihood of responding to treatment with the IL-17 antagonist, and the absence of the at least one dual gene indicates that the patient will be against the IL -17 antagonist treatment is less likely to respond. A method of predicting the likelihood that a patient with RA will respond to treatment with an IL-17 antagonist, comprising analyzing the presence or absence of SE in a biological sample from the patient using an automated analyzer, wherein the SE indication is present The patient will have an increased likelihood of responding to treatment with the IL-17 antagonist, and the absence of the SE indicates that the patient will be less likely to respond to treatment with the IL-17 antagonist. The method of any one of claims 29 or 30, wherein the presence or absence of the SE is detected by analyzing the presence or absence of at least one of the dual genes in the HLA-DRB1*SE dual genome in the biological sample. The method, use or kit of any of the above claims, wherein the SE, the at least one dual gene in the HLA-DRB1*04 dual genome or the at least one dual in the HLA-DRB1*SE dual genome The presence or absence of a gene is detected by a technique selected from the group consisting of Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR), RT-PCR ELISA, TaqMan-based quantitative RT-PCR. (Probe-based quantitative RT-PCR), SYBR-based quantitative RT-PCR, polymerase chain reaction (PCR), direct sequencing, sequence-specific oligonucleotide (SSO) hybridization, sequence-specific primer (SSP) ) typing, and sequence-based typing (SBT), Southern dot method, quantitative PCR (probe based or based on SYBR green), immunoassay, immunohistochemistry, ELISA, flow cytometry, Western blots Method, HPLC and mass spectrometry. The method, use, or kit of any of the preceding claims, wherein the biological sample is selected from the group consisting of: synovial fluid, blood, serum, plasma, urine, tears, saliva, cerebrospinal fluid, white blood cell samples, and tissue sample. The method, use, or kit of any of the preceding claims, wherein the patient is a high risk RA patient. A kit comprising: a) a pharmaceutical composition comprising an IL-17 antagonist for treating rheumatoid arthritis (RA) in a patient; and b) instructions describing how to administer the pharmaceutical composition to the patient, The patient is characterized by having a dual gene in the SE, HLA-DRB1*04 dual genome or a dual gene in the HLA-DRB1*SE dual genome. Use of an IL-17 antagonist for the preparation of a medicament for the treatment of RA, the restriction being that the patient is based on a dual gene in the SE, HLA-DRB1*04 dual genome or a duality in the HLA-DRB1*SE dual genome The gene is selected for treatment. The method, use, or kit of any of the preceding claims, wherein the patient's RA has not previously been treated with a TNFα antagonist. The method, use or kit of any of the above claims, wherein the IL-17 antagonist is an IL-17 binding molecule or an IL-17 receptor binding molecule. The method, use or kit of claim 38, wherein the IL-17 binding molecule or IL-17 receptor binding molecule is an IL-17 binding molecule selected from the group consisting of: a) secucumumab (secukinumab b) binds to IL-17 antibody comprising the following epitopes in IL-17: Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129; c) binding to IL- An IL-17 antibody comprising the following epitopes: Tyr43, Tyr44, Arg46, Ala79, Asp80; d) binding to an epitope of an IL-17 homodimer having two mature IL-17 protein chains An IL-17 antibody comprising Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129 in one strand and Tyr43, Tyr44, Arg46, Ala79 in another chain, Asp80; e) an IL-17 antibody that binds to an epitope of an IL-17 homodimer of two mature IL-17 protein chains, which comprises Leu74, Tyr85, His86, Met87 in one strand, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129 and on another chain Including Tyr43, Tyr44, Arg46, Ala79, Asp80, wherein the IL-17 binding K _D molecule is from about 100 pM to 200 pM, and wherein the IL-17 binding activity of the molecule in vivo half-life of about 4 weeks; and f) comprising IL-17 antibody is selected from the group consisting of: i) comprising the SEQ ID NO: 8 shown in an immunoglobulin heavy chain variable domain amino acid sequence (V _H); ii) comprising the SEQ ID NO: 10 shown in an immunoglobulin light chain variable domain amino acid sequence (V _L); iii) as containing the SEQ ID NO: 8 shown in immunoglobulin V _H domain comprises the amino acid sequence and the as SEQ ID NO: 10 shown in immunoglobulin V _L domain of the amino acid sequence; IV) comprising an immunoglobulin V _H domain as shown in the following variations of high regions: SEQ ID NO: 1, SEQ ID NO : 2 and SEQ ID NO: 3; v) comprising an immunoglobulin V _L domain as shown in the following variations of high regions: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; vi) comprising the The immunoglobulin _VH domain of the hypervariable region shown below: SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13; vii) immunoglobulins comprising the hypervariable regions shown below V _H domain: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, and comprising The other hypervariable regions of immunoglobulin V shown of the _L domain: SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and viii) those comprising the following hypervariable region as shown in the immunoglobulin V _H domain: SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, and the immunoglobulin V _L domain comprises hypervariable regions such as shown in the following of: SEQ ID NO: 4 SEQ ID NO: 5 and SEQ ID NO: 6. The method, use, or kit of claim 39, wherein the IL-17 binding molecule is cesikumab.