Use of pad inhibitor for treatment or prevention of autoimmune arthritis

Abstract

The present invention provides drugs for treating or preventing autoimmune arthritis. The drugs are 1) a drug that contains a peptidylarginine deiminase inhibitor as an active ingredient and is for administration to subjects who do not respond to at least one disease-modifying antirheumatic drug or subjects who are predicted to not respond to disease-modifying antirheumatic drugs and 2) a drug that includes a peptidylarginine deiminase inhibitor and a tumor necrosis factor inhibitor.

Description

Use of a PAD inhibitor for the treatment or prevention of autoimmune arthritis

[0001] Reference to the Sequence ListingThis specification refers to the sequence listing electronically submitted as an.xml file named "P251302WO". This.xml file was generated on October 24, 2025 and is 61 kilobytes in size. The entire content of the sequence listing is incorporated herein by reference.

[0002] The present disclosure relates to the use of a peptidylarginine deiminase (PAD) inhibitor for the treatment or prevention of autoimmune arthritis. The present disclosure also relates to a biomarker indicating the effectiveness of treating or preventing autoimmune arthritis and a method for predicting the effectiveness of treating or preventing autoimmune arthritis using the biomarker.

[0003] Disease-modifying antirheumatic drugs such as tumor necrosis factor (TNF) inhibitors are used for the treatment of autoimmune arthritis. However, there are many patients who do not respond to these drugs, and there is a need for a more effective therapeutic agent for autoimmune arthritis that can also be applied to such patients. On the other hand, Patent Document 1 discloses an anti-PAD4 antibody. The entire content of this published gazette is incorporated herein by reference.

[0004] International Publication No. 2022 / 176970

[0005] The present disclosure provides a more effective therapeutic agent for autoimmune arthritis that can also be applied to patients who do not respond to disease-modifying antirheumatic drugs. The present disclosure also provides a biomarker for predicting the effectiveness of treating or preventing autoimmune arthritis.

[0006] The disclosure includes the following embodiments: [1] A pharmaceutical for the treatment or prevention of autoimmune arthritis, comprising a peptidylarginine deiminase (PAD) inhibitor as an active ingredient, and administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. [1] can be rephrased as [1'], [1'-1], [1''], [1'''], [1'''']. [1'] A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient, and administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. [1'-1] A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient and a pharmacologically acceptable carrier, to be administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. [1''] A method for the treatment or prevention of autoimmune arthritis, comprising administering an effective amount of a PAD inhibitor to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. [1''] The use of a PAD inhibitor in the manufacture of a medicament for the treatment or prevention of autoimmune arthritis, wherein the medicament is administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. [1''''] A PAD inhibitor for use in the treatment or prevention of autoimmune arthritis, which is administered to subjects unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug.

[0007] The specific embodiments of [1] below are also applicable to [1'], [1''], [1'''], and [1'''']. [2] The pharmaceutical product according to [1], wherein the subject predicted to be unresponsive to the disease-modifying antirheumatic drug is a subject whose blood level of neutrophil activation-related factor is higher than the reference value. [3] The pharmaceutical product according to [2], wherein the reference value is the level of neutrophil activation-related factor in the blood of a healthy person or a patient with low disease activity of autoimmune arthritis. [4] The pharmaceutical product according to any one of [1] to [3], wherein the disease-modifying antirheumatic drug is a synthetic antirheumatic drug or a biological antirheumatic drug. [5] The pharmaceutical product according to [4], wherein the synthetic antirheumatic drug is a conventional synthetic antirheumatic drug or a molecularly targeted synthetic antirheumatic drug. [6] The pharmaceutical product according to [5], wherein the conventional synthetic antirheumatic drug is methotrexate. [7] The pharmaceutical product according to [5], wherein the molecularly targeted synthetic antirheumatic drug is a janus kinase inhibitor. [8] The pharmaceutical product according to [4], wherein the biological antirheumatic drug is a tumor necrosis factor (TNF) inhibitor, an interleukin (IL)-6 inhibitor, or a T cell selective costimulation modifier. [9] The pharmaceutical product according to [8], wherein the TNF inhibitor is an anti-TNF antibody or a TNF-alpha / lymphotoxin-alpha receptor-Fc fusion protein.

[10] The pharmaceutical product according to [8], wherein the IL-6 inhibitor is an anti-IL-6 receptor antibody.

[11] The pharmaceutical product according to [8], wherein the T cell selective costimulation modifier is a cytotoxic T-lymphocyte antigen-4-Fc fusion protein.

[12] The pharmaceutical product according to any one of [1] to

[11] , wherein the autoimmune arthritis is rheumatoid arthritis.

[13] The pharmaceutical product according to any one of [1] to

[12] , wherein the PAD inhibitor is an anti-PAD antibody or a fragment of that antibody.

[0008]

[14] The anti-PAD antibody or antibody fragment thereof is: a-1) an anti-PAD4 antibody or antibody fragment thereof in which the complementarity determining region (CDR) 1 of the heavy chain contains the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain contains the amino acid sequence of SEQ ID NO: 7, the CDR3 of the heavy chain contains the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain contains the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain contains the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain contains the amino acid sequence of SEQ ID NO: 13; b-1) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain contains the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain contains the amino acid sequence of SEQ ID NO: 8, the CDR3 of the heavy chain contains the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain contains the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain contains the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain contains the amino acid sequence of SEQ ID NO: 13. c-1) An anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 14; and d-1) An anti-PAD4 antibody or an antibody fragment thereof, which is selected from the above.

[0009]

[15] The anti-PAD antibody or antibody fragment thereof is: a-2) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 7, the CDR3 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain consists of the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain consists of the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain consists of the amino acid sequence of SEQ ID NO: 13; b-2) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 8, the CDR3 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain consists of the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain consists of the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain consists of the amino acid sequence of SEQ ID NO: 13; c-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 14; and d-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 15.

[0010]

[16] The anti-PAD antibody or antibody fragment thereof is: a-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23. A pharmaceutical product according to

[13] , selected from the group consisting of the following.

[0011]

[17] The anti-PAD antibody or antibody fragment thereof is: a-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-4) A pharmaceutical product according to

[13] or

[16] , selected from the group consisting of an anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23.

[0012]

[18] A pharmaceutical for the treatment or prevention of autoimmune arthritis, comprising a peptidylarginine deiminase (PAD) inhibitor and a tumor necrosis factor (TNF) inhibitor.

[18] may be replaced with [18'], [18''], [18'''], or [18'''']. [18'] A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor and a TNF inhibitor as active ingredients. [18''] A method for the treatment or prevention of autoimmune arthritis, comprising administering an effective amount of a PAD inhibitor and a TNF inhibitor to a subject in need thereof. [18'''] Use of a PAD inhibitor and a TNF inhibitor in the manufacture of a pharmaceutical for the treatment or prevention of autoimmune arthritis. [18''''] A combination of a PAD inhibitor and a TNF inhibitor for use in the treatment or prevention of autoimmune arthritis.

[0013] The following specific embodiments of

[18] are also incorporated into [18'], [18''], [18'''], and [18''''].

[19] The pharmaceutical product according to

[18] , wherein the autoimmune arthritis is rheumatoid arthritis.

[20] The pharmaceutical product according to

[18] , wherein the PAD inhibitor is an anti-PAD antibody or a fragment of that antibody.

[21] The anti-PAD antibody or antibody fragment thereof is: a-1) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 contains the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 contains the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR3 contains the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 contains the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 contains the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 contains the amino acid sequence of SEQ ID NO: 13; b-1) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 contains the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 contains the amino acid sequence of SEQ ID NO: 8, the heavy chain CDR3 contains the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 contains the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 contains the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 contains the amino acid sequence of SEQ ID NO: 13. c-1) an anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: a heavy chain CDR1 containing the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR2 containing the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR3 containing the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 containing the amino acid sequence of SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence of SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence of SEQ ID NO: 14; and d-1) an anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: a heavy chain CDR1 containing the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR2 containing the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR3 containing the amino acid sequence of SEQ ID NO: 11, a light chain CDR1 containing the amino acid sequence of SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence of SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence of SEQ ID NO: 15.

[0014]

[22] The anti-PAD antibody or antibody fragment thereof is: a-2) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 7, the CDR3 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain consists of the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain consists of the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain consists of the amino acid sequence of SEQ ID NO: 13; b-2) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 8, the CDR3 of the heavy chain consists of the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain consists of the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain consists of the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain consists of the amino acid sequence of SEQ ID NO: 13; c-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 14; and d-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 15.

[0015]

[23] The anti-PAD antibody or antibody fragment thereof is: a-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23. A pharmaceutical product selected from the group consisting of

[20] .

[0016]

[24] The anti-PAD antibody or antibody fragment thereof is: a-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-4) A pharmaceutical product according to

[20] or

[23] , selected from the group consisting of an anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23.

[25] A pharmaceutical product according to any one of

[18] to

[24] , wherein the TNF inhibitor is a composition comprising an anti-TNF antibody or a TNF-alpha / lymphotoxin-alpha receptor-Fc fusion protein.

[26] A pharmaceutical product according to any one of

[18] to

[25] , administered to a subject unresponsive to at least one disease-modifying antirheumatic drug or a subject predicted to be unresponsive to a disease-modifying antirheumatic drug.

[27] A pharmaceutical product according to

[26] , wherein the subject predicted to be unresponsive to the disease-modifying antirheumatic drug has a blood level of neutrophil activation-related factor higher than the reference value.

[28] The pharmaceutical product according to

[27] , wherein the reference value is the level of neutrophil activation-related factor in the blood of a healthy person or a patient with low disease activity of autoimmune arthritis.

[29] The pharmaceutical product according to any one of

[26] to

[28] , wherein the disease-modifying antirheumatic drug is a synthetic antirheumatic drug or a biological antirheumatic drug.

[30] The pharmaceutical product according to

[29] , wherein the synthetic antirheumatic drug is a conventional synthetic antirheumatic drug or a molecularly targeted synthetic antirheumatic drug.

[31] The pharmaceutical product according to

[30] , wherein the conventional synthetic antirheumatic drug is methotrexate.

[32] The pharmaceutical product according to

[30] , wherein the molecularly targeted synthetic antirheumatic drug is a janus kinase inhibitor.

[33] The pharmaceutical product according to

[29] , wherein the biological antirheumatic drug is a tumor necrosis factor (TNF) inhibitor, an interleukin (IL)-6 inhibitor, or a T cell selective costimulation modifier.

[34] The pharmaceutical product according to

[33] , wherein the TNF inhibitor is an anti-TNF antibody or a TNF-alpha / lymphotoxin-alpha receptor-Fc fusion protein.

[35] The pharmaceutical product according to

[33] , wherein the IL-6 inhibitor is an anti-IL-6 receptor antibody.

[36] The pharmaceutical product according to

[33] , wherein the T cell selective costimulation modifier is a cytotoxic T-lymphocyte antigen-4-Fc fusion protein.

[0017] One embodiment of the present disclosure is a pharmaceutical product for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient, which is administered to a subject whose blood level of neutrophil activation-related factor is higher than a reference value. In other words, the use of a PAD inhibitor in the manufacture of a pharmaceutical product for the treatment or prevention of autoimmune arthritis, wherein the pharmaceutical product is administered to a subject whose blood level of neutrophil activation-related factor is higher than a reference value; a PAD inhibitor for use in the treatment or prevention of autoimmune arthritis, wherein an effective amount of the PAD inhibitor is administered to a subject whose blood level of neutrophil activation-related factor is higher than a reference value; and a method for the treatment or prevention of autoimmune arthritis, comprising: measuring or receiving the measurement result of the level of neutrophil activation-related factor in the blood of a subject; comparing the level of the neutrophil activation-related factor with a reference value; selecting a subject if the level of the neutrophil activation-related factor is higher than a reference value; and administering an effective amount of the PAD inhibitor to the selected subject.

[0018] Another embodiment of the present disclosure is a method for predicting the efficacy of a composition for the treatment or prevention of autoimmune arthritis comprising a PAD inhibitor, comprising: measuring or receiving the level of neutrophil activation-related factor in the blood of a subject before administration of the composition; measuring or receiving the level of neutrophil activation-related factor in the blood of a subject after administration of the composition; comparing the level of neutrophil activation-related factor measured before administration of the composition with the level of neutrophil activation-related factor measured after administration of the composition; and predicting that the composition is effective in treating or preventing the subject if the level of neutrophil activation-related factor measured after administration of the composition is lower than the level of neutrophil activation-related factor measured before administration of the composition. A further embodiment of the present disclosure is a method for predicting the efficacy of a composition for the treatment or prevention of autoimmune arthritis comprising a PAD inhibitor, comprising: measuring or receiving the level of neutrophil activation-related factor in the blood of a subject before administration of the composition; administering the composition to the subject; measuring or receiving the level of neutrophil activation-related factor in the blood of the subject after administration of the composition; comparing the level of neutrophil activation-related factor measured before administration of the composition with the level of neutrophil activation-related factor measured after administration of the composition; and predicting that the composition is effective in treating or preventing the subject if the level of neutrophil activation-related factor measured after administration of the composition is lower than the level of neutrophil activation-related factor measured before administration of the composition.

[0019] This disclosure provides a more effective treatment for autoimmune arthritis that can be applied to patients who are unresponsive to disease-modifying antirheumatic drugs. Furthermore, by measuring the amount of biomarkers, the effectiveness of treating autoimmune arthritis with PAD inhibitors can be predicted, leading to more effective treatment.

[0020] Figure 1 is a graph comparing PAD4 protein concentration and anti-citrullinated peptide antibody (ACPA) concentration in each group: healthy individuals (HC), patients responding to TNF inhibitors (TNF-R), patients not responding to TNF inhibitors (TNF-IR), and patients not responding to biological anti-rheumatic drugs other than TNF inhibitors (non-TNF-IR). "pre" indicates before administration of each drug, and "post" indicates after administration of each drug. Figure 2 is a graph showing arthritis scores, citrullinated protein levels, CXC motif chemokine ligand 13 (CXCL13) protein concentration, and ACPA concentration after administration of various drugs in collagen-induced arthritis (CIA) model mice. "Normal" indicates normal mice and untreated mice. Figure 3 is a graph showing arthritis scores in etanercept-unresponsive CIA model mice with continued etanercept administration or anti-PAD4 antibody administration. Figure 4 is a graph showing the time course of arthritis scores in CIA model mice under control, etanercept administration, anti-PAD4 antibody administration, and combination administration of these. Figure 5 is a photographic diagram showing the results of PAD4 immunostaining in the joints of CIA model mice.

[0021] <1. Embodiments in which a PAD inhibitor is administered to subjects unresponsive to or expected to be unresponsive to disease-modifying antirheumatic drugs> One embodiment of the present disclosure is a pharmaceutical for the treatment or prevention of autoimmune arthritis, comprising a peptidylarginine deiminase (PAD) inhibitor as an active ingredient, and administered to subjects unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. This embodiment can be rephrased as follows: A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient, and administered to subjects unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. A method for the treatment or prevention of autoimmune arthritis, comprising administering an effective amount of a PAD inhibitor to subjects unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. The use of a PAD inhibitor in the manufacture of a pharmaceutical product for the treatment or prevention of autoimmune arthritis, wherein the pharmaceutical product is administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug. A PAD inhibitor for use in the treatment or prevention of autoimmune arthritis, wherein the PAD inhibitor is administered to a subject unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug.

[0022] In this disclosure, treatment includes any treatment of a disease of a mammal, in particular a human, and includes inhibiting disease symptoms, i.e., stopping their progression or eliminating the disease or symptoms, and reducing disease symptoms, i.e., causing a regression of the disease or symptoms or delaying the progression of the symptoms.

[0023] In this disclosure, prevention includes delaying or preventing the onset or occurrence of the disease in mammals, particularly humans. Prevention includes relapse prevention, where relapse prevention includes preventing relapses of the disease in mammals, particularly humans, that involve repeated remissions and relapses. Prevention also includes reducing the risk of developing the disease or its incidence or relapse rate.

[0024] Autoimmune arthritis is, for example, rheumatoid arthritis.

[0025] PAD inhibitors are administered to subjects who are unresponsive to at least one disease-modifying antirheumatic drug or who are expected to be unresponsive to disease-modifying antirheumatic drugs. Pharmaceuticals containing PAD inhibitors may have a statement in their package insert to the effect that they are "administered to subjects who are unresponsive to disease-modifying antirheumatic drugs or who are expected to be unresponsive to disease-modifying antirheumatic drugs."

[0026] In this disclosure, the subject (sometimes referred to as the patient) includes one or more non-human mammals, specifically, for example, mice, guinea pigs, hamsters, rats, rodents, rabbits, pigs, sheep, goats, cattle, horses, cats, dogs, marmosets, monkeys, or chimpanzees.

[0027] Disease-modifying antirheumatic drugs (DMARDs) are, for example, synthetic disease-modifying antirheumatic drugs (sDMARDs) or biological disease-modifying antirheumatic drugs (bDMARDs). Synthetic disease-modifying antirheumatic drugs are, for example, conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) or targeted synthetic disease-modifying antirheumatic drugs (tsDMARDs). An example of a conventional synthetic disease-modifying antirheumatic drug is methotrexate. An example of a targeted synthetic disease-modifying antirheumatic drug is a janus kinase (JAK) inhibitor, more specifically tofacitinib. Biological antirheumatic drugs include, for example, tumor necrosis factor (TNF) inhibitors, interleukin-6 (IL-6) inhibitors, or T cell selective costimulation modulogenators. TNF inhibitors suppress the action of TNF in vivo and are either anti-TNF antibodies or tumor necrosis factor-alpha / lymphotoxin-alpha (TNFα / LTα) receptor-Fc fusion proteins, more specifically infliximab, etanercept, adalimumab, certolizumab, pegol, and golimumab. Anti-TNF antibodies are preferably anti-TNF-α antibodies. IL-6 inhibitors suppress the action of IL-6 in vivo and inhibit T cell activation by inhibiting costimulation signals between antigen-presenting cells and T cells, for example, anti-IL-6 receptor antibodies, more specifically tocilizumab.The T-cell selective costimulatory modulo agent is, for example, a cytotoxic T-lymphocyte antigen-4 (CTLA-4)-Fc fusion protein, more specifically abatacept.

[0028] The subjects may, for example, be unresponsive to at least one, at least two, at least three, at least four, or all disease-modifying antirheumatic drugs.

[0029] In this disclosure, non-response refers to, for example, a case where disease activity is assessed using the rheumatoid arthritis disease activity assessment method within six months of starting administration of a disease-modifying antirheumatic drug, and disease activity does not improve within three months, or where clinical remission or low disease activity is not achieved within six months. This can be determined using, for example, the DAS28-ESR, SDAI, and CDAI scores. For example, if autoimmune arthritis is rheumatoid arthritis, it refers to a case where no improvement in the DAS28-ESR, SDAI, or CDAI scores is observed during a treatment period of about three months. Alternatively, it refers to a case where the DAS28-ESR remains above 3.2, the SDAI above 11, or the CDAI above 10 during a treatment period of about six months.

[0030] The aforementioned "DAS28-ESR" refers to an index for assessing disease activity in rheumatoid arthritis (Reference: Arthritis Rheum. 1995 Jan;38(1):44-8. PMID: 7818570), and is an abbreviation for Disease Activity Score-28 for Rheumatoid Arthritis with Erythrocyte Sedimentation Rate. DAS28-ESR can be calculated using the following formula (1): Formula (1): DAS28-ESR = 0.56 × √Number of tender joints + 0.28 × √Number of swollen joints + 0.7 × LN (ESR) + 0.014 × VAS Depending on the value of DAS28-ESR, disease activity can be evaluated as follows. DAS28-ESR < 2.6: Remission 2.6 ≤ DAS28-ESR < 3.2: Low disease activity 3.2 ≤ DAS28-ESR ≤ 5.1: Moderate disease activity 5.1 < DAS28-ESR: High disease activity

[0031] The aforementioned "SDAI" is an abbreviation for Simplified Disease Activity Index (Reference: Rheumatology. 2003 Feb;42(2):244-57. PMID: 12595618), and is calculated from five elements: number of tender joints (TJC28), number of swollen joints (SJC28), overall assessment by the patient, overall assessment by the physician, and CRP value. SDAI ≤ 3.3: Remission 3.3 < SDAI ≤ 11: Low disease activity 11 < SDAI ≤ 26: Moderate disease activity 26 < SDAI: High disease activity

[0032] The aforementioned "CDAI" is an abbreviation for Clinical Disease Activity Index (Reference: Arthritis Res Ther. 2005;7(4):R796-806.PMID:15987481), and is calculated from four elements: number of tender joints (TJC28), number of swollen joints (SJC28), overall assessment by the patient, and overall assessment by the physician. CDAI ≤ 2.8: Remission 2.8 < CDAI ≤ 10: Low disease activity 10 < CDAI ≤ 22: Moderate disease activity 22 < CDAI: High disease activity

[0033] PAD inhibitors may be administered to patients predicted to be unresponsive to disease-modifying antirheumatic drugs. Patients predicted to be unresponsive to disease-modifying antirheumatic drugs may be those whose levels of neutrophil activation-related factors in the blood are higher than the reference range. Here, neutrophil activation-related factors are selected from the group consisting of, for example, PAD protein, myeloperoxidase (MPO) protein, CXCL13 protein, citrullinated proteins, and ACPA. PAD may also be PAD4. These measurement methods, as well as "reference range" and "higher than reference range," will be described later.

[0034] PAD inhibitors include, for example, PAD4 inhibitors. PAD inhibitors are, for example, anti-PAD antibodies or their antibody fragments, and anti-PAD4 antibodies or their antibody fragments. An anti-PAD antibody refers to an immunoglobulin molecule or a modified thereof that binds to PAD. Modified molecules include multispecific antibodies, chimeric antibodies, humanized antibodies, functionally modified antibodies, and conjugate antibodies.

[0035] A multispecific antibody is an asymmetric antibody that possesses two or more independent antigen recognition sites with two or more different antigen specificities. Examples include bispecific antibodies with two antigen specificities and trispecific antibodies with three antigen specificities. An example of a multispecific antibody is an anti-PAD2 / PAD4 bispecific antibody.

[0036] A chimeric antibody is an antibody in which the light chain, heavy chain, or both consist of a variable region of non-human origin and a constant region of human origin.

[0037] A humanized antibody is an antibody that consists of a variable region comprising a complementarity-determining region of a non-human animal-derived antibody and a framework region derived from a human antibody, and a constant region derived from a human antibody.

[0038] In this disclosure, a functionally modified antibody refers to an antibody whose functions other than antigen-binding function, such as cell-killing function, complement-activating function, or blood half-life extension function, have been modified by altering the antibody sequence, sugar chain, etc.

[0039] In this disclosure, a conjugated antibody refers to an antibody to which a functional molecule other than the antibody, such as a non-peptide polymer like polyethylene glycol (PEG), a radioactive substance, a toxin, a small molecule compound, a cytokine, a growth factor, albumin, or an enzyme, has been chemically or genetically engineered to it.

[0040] In this disclosure, unless otherwise specified, "antibody fragment" means "antigen-binding fragment." Antibody fragments may also be called antigen-binding molecules. An antigen-binding fragment is a protein containing a portion of an antibody that can bind to an antigen. Examples of antigen-binding fragments include F(ab')2, Fab', Fab, Fv (variable fragment of antibody), disulfide-bonded Fv, single-chain antibodies (scFv), and polymers thereof. Furthermore, antigen-binding fragments include conjugated antigen-binding fragments to which functional molecules such as non-peptide polymers like polyethylene glycol (PEG), radioactive materials, toxins, small molecule compounds, cytokines, growth factors (TGF-β, NGF, Neurotrophin, etc.), albumin, enzymes, and other antibodies are chemically or genetically engineered to be bound.

[0041] Detailed information on the amino acid sequences of PADs can be found on websites such as NCBI (National Center for Biotechnology Information) or HGNC (HUGO Gene Nomenclature Committee). For example, the amino acid sequence of PAD4 is sequence number 45. PAD4 can be derived from any organism as long as it possesses PAD4 activity. For example, the amino acid sequence of PAD2 is sequence number 46. PAD2 can be derived from any organism as long as it possesses PAD2 activity.

[0042] Anti-PAD antibodies can be antibodies that possess heavy and light chains and have specific heavy chain complementarity-determining region sequences and light chain complementarity-determining region sequences. Complementarity-determining regions (CDRs) are the variable regions of an immunoglobulin molecule that form the antigen-binding site. Also known as hypervariable regions, they are the parts of the immunoglobulin molecule where amino acid sequence changes are particularly significant. There are three CDRs in both the light and heavy chains (LCDR1, LCDR2, LCDR3, and HCDR1, HCDR2, HCDR3). The CDRs of immunoglobulin molecules can be determined according to the Kabat numbering system, the IMGT method, the Chothia method, or the AbM method. Sequence IDs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 13, 14, 15, 24, 25, 26, 29, 30, 31, 32, 33, 34, 38, 39, 40, 41, and 42 in this specification are determined by the Kabat method, sequence IDs 11 and 12 are determined by the IMGT method, and sequence ID 37 is determined by the AbM method.

[0043] An anti-PAD4 antibody or antibody fragment thereof may be, for example, a-1) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 (HCDR1) contains the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 (HCDR2) contains the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR3 (HCDR3) contains the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 (LCDR1) contains the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 (LCDR2) contains the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 (LCDR3) contains the amino acid sequence of SEQ ID NO: 13; b-1) an anti-PAD4 antibody or antibody fragment thereof in which HCDR1 contains the amino acid sequence of SEQ ID NO: 1, HCDR2 contains the amino acid sequence of SEQ ID NO: 8, HCDR3 contains the amino acid sequence of SEQ ID NO: 11, LCDR1 contains the amino acid sequence of SEQ ID NO: 4, LCDR2 contains the amino acid sequence of SEQ ID NO: 5, and LCDR3 contains the amino acid sequence of SEQ ID NO: 13. c-1) An anti-PAD4 antibody or an antibody fragment thereof, wherein HCDR1 contains the amino acid sequence of SEQ ID NO: 1, HCDR2 contains the amino acid sequence of SEQ ID NO: 9, HCDR3 contains the amino acid sequence of SEQ ID NO: 12, LCDR1 contains the amino acid sequence of SEQ ID NO: 4, LCDR2 contains the amino acid sequence of SEQ ID NO: 5, and LCDR3 contains the amino acid sequence of SEQ ID NO: 14; and d-1) An anti-PAD4 antibody or an antibody fragment thereof, wherein HCDR1 contains the amino acid sequence of SEQ ID NO: 1, HCDR2 contains the amino acid sequence of SEQ ID NO: 10, HCDR3 contains the amino acid sequence of SEQ ID NO: 11, LCDR1 contains the amino acid sequence of SEQ ID NO: 4, LCDR2 contains the amino acid sequence of SEQ ID NO: 5, and LCDR3 contains the amino acid sequence of SEQ ID NO: 15. e-1) An anti-PAD4 antibody or an antibody fragment thereof, wherein HCDR1 contains the amino acid sequence of SEQ ID NO: 29, HCDR2 contains the amino acid sequence of SEQ ID NO: 30, HCDR3 contains the amino acid sequence of SEQ ID NO: 31, LCDR1 contains the amino acid sequence of SEQ ID NO: 32, LCDR2 contains the amino acid sequence of SEQ ID NO: 33, and LCDR3 contains the amino acid sequence of SEQ ID NO: 34, andf-1) Selected from the group consisting of an anti-PAD4 antibody or an antibody fragment thereof, wherein HCDR1 contains the amino acid sequence of SEQ ID NO: 37, HCDR2 contains the amino acid sequence of SEQ ID NO: 38, HCDR3 contains the amino acid sequence of SEQ ID NO: 39, LCDR1 contains the amino acid sequence of SEQ ID NO: 40, LCDR2 contains the amino acid sequence of SEQ ID NO: 41, and LCDR3 contains the amino acid sequence of SEQ ID NO: 42.

[0044] Alternatively, an anti-PAD4 antibody or antibody fragment thereof may be, for example, a-2) an anti-PAD4 antibody or antibody fragment thereof in which HCDR1 consists of the amino acid sequence of SEQ ID NO: 1, HCDR2 consists of the amino acid sequence of SEQ ID NO: 7, HCDR3 consists of the amino acid sequence of SEQ ID NO: 11, LCDR1 consists of the amino acid sequence of SEQ ID NO: 4, LCDR2 consists of the amino acid sequence of SEQ ID NO: 5, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 13; b-2) an anti-PAD4 antibody or antibody fragment thereof in which HCDR1 consists of the amino acid sequence of SEQ ID NO: 1, HCDR2 consists of the amino acid sequence of SEQ ID NO: 8, HCDR3 consists of the amino acid sequence of SEQ ID NO: 11, LCDR1 consists of the amino acid sequence of SEQ ID NO: 4, LCDR2 consists of the amino acid sequence of SEQ ID NO: 5, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 13. c-2) An anti-PAD4 antibody or antibody fragment thereof, wherein HCDR1 consists of the amino acid sequence of SEQ ID NO: 1, HCDR2 consists of the amino acid sequence of SEQ ID NO: 9, HCDR3 consists of the amino acid sequence of SEQ ID NO: 12, LCDR1 consists of the amino acid sequence of SEQ ID NO: 4, LCDR2 consists of the amino acid sequence of SEQ ID NO: 5, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 14; and d-2) An anti-PAD4 antibody or antibody fragment thereof, wherein HCDR1 consists of the amino acid sequence of SEQ ID NO: 1, HCDR2 consists of the amino acid sequence of SEQ ID NO: 10, HCDR3 consists of the amino acid sequence of SEQ ID NO: 11, LCDR1 consists of the amino acid sequence of SEQ ID NO: 4, LCDR2 consists of the amino acid sequence of SEQ ID NO: 5, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 15. e-2) An anti-PAD4 antibody or an antibody fragment thereof, wherein HCDR1 consists of the amino acid sequence of SEQ ID NO: 29, HCDR2 consists of the amino acid sequence of SEQ ID NO: 30, HCDR3 consists of the amino acid sequence of SEQ ID NO: 31, LCDR1 consists of the amino acid sequence of SEQ ID NO: 32, LCDR2 consists of the amino acid sequence of SEQ ID NO: 33, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 34, and(f-2) The anti-PAD4 antibody or its antibody fragment is selected from the group consisting of an anti-PAD4 antibody or its antibody fragment in which HCDR1 consists of the amino acid sequence of SEQ ID NO: 37, HCDR2 consists of the amino acid sequence of SEQ ID NO: 38, HCDR3 consists of the amino acid sequence of SEQ ID NO: 39, LCDR1 consists of the amino acid sequence of SEQ ID NO: 40, LCDR2 consists of the amino acid sequence of SEQ ID NO: 41, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 42.

[0045] Alternatively, the anti-PAD4 antibody or its antibody fragment is, for example, (a-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of amino acid numbers 1 to 120 of SEQ ID NO: 16 and the light chain variable region contains the amino acid sequence of amino acid numbers 1 to 105 of SEQ ID NO: 17; (b-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of amino acid numbers 1 to 120 of SEQ ID NO: 18 and the light chain variable region contains the amino acid sequence of amino acid numbers 1 to 105 of SEQ ID NO: 19; (c-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of amino acid numbers 1 to 120 of SEQ ID NO: 20 and the light chain variable region contains the amino acid sequence of amino acid numbers 1 to 105 of SEQ ID NO: 21; (d-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of amino acid numbers 1 to 120 of SEQ ID NO: 22 and the light chain variable region contains the amino acid sequence of amino acid numbers 1 to 105 of SEQ ID NO: 23; (e-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 35 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 36; and (f-3) an anti-PAD4 antibody or its antibody fragment in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 43 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 44, and is selected from the group consisting of these.

[0046] Alternatively, an anti-PAD4 antibody or antibody fragment thereof may be, for example, a-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21. d-4) An anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23; e-4) An anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 35 and the light chain variable region consists of the amino acid sequence of SEQ ID NO: 36; and f-4) An anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 43 and the light chain variable region consists of the amino acid sequence of SEQ ID NO: 44.

[0047] Alternatively, the anti-PAD4 antibody or antibody fragment thereof is selected from the group consisting of: a-5) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain consists of the amino acid sequence of SEQ ID NO: 16 and the light chain consists of the amino acid sequence of SEQ ID NO: 17; b-5) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain consists of the amino acid sequence of SEQ ID NO: 18 and the light chain consists of the amino acid sequence of SEQ ID NO: 19; c-5) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain consists of the amino acid sequence of SEQ ID NO: 20 and the light chain consists of the amino acid sequence of SEQ ID NO: 21; and d-5) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain consists of the amino acid sequence of SEQ ID NO: 22 and the light chain consists of the amino acid sequence of SEQ ID NO: 23.

[0048] The anti-PAD2 antibody or an antibody fragment thereof is selected from the group consisting of, for example, g-1) an anti-PAD4 antibody or an antibody fragment thereof in which HCDR1 contains the amino acid sequence of SEQ ID NO: 2, HCDR2 contains the amino acid sequence of SEQ ID NO: 3, HCDR3 contains the amino acid sequence of SEQ ID NO: 6, LCDR1 contains the amino acid sequence of SEQ ID NO: 24, LCDR2 contains the amino acid sequence of SEQ ID NO: 25, and LCDR3 contains the amino acid sequence of SEQ ID NO: 26, and g-2) an anti-PAD4 antibody or an antibody fragment thereof in which HCDR1 consists of the amino acid sequence of SEQ ID NO: 2, HCDR2 consists of the amino acid sequence of SEQ ID NO: 3, HCDR3 consists of the amino acid sequence of SEQ ID NO: 6, LCDR1 consists of the amino acid sequence of SEQ ID NO: 24, LCDR2 consists of the amino acid sequence of SEQ ID NO: 25, and LCDR3 consists of the amino acid sequence of SEQ ID NO: 26.

[0049] Alternatively, the anti-PAD2 antibody or an antibody fragment thereof is selected from the group consisting of, for example, g-3) an anti-PAD4 antibody or an antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 27 and the light chain variable region contains the amino acid sequence of SEQ ID NO: 28, and g-4) an anti-PAD4 antibody or an antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 27 and the light chain variable region consists of the amino acid sequence of SEQ ID NO: 28.

[0050] Alternatively, the anti-PAD4 antibody, anti-PAD2 / PAD4 bispecific antibody or an antibody fragment thereof can incorporate, for example, the contents described in International Publication No. 2022 / 176970, International Publication No. 2024 / 020579, and International Publication No. 2024 / 133161, and can be produced by the methods described in these documents.

[0051] In addition, in the heavy chain variable region and / or light chain variable region of the amino acid sequence of the anti-PAD4 antibody, one or more amino acids (1 to 20, 1 to 10, or 1 to 5) may be substituted, deleted, added, or inserted, as long as the ability to bind to PAD4 (preferably with a KD value of 100 pM or less to PAD4, more preferably 90 pM or less, and even more preferably 80 pM or less) is maintained and the property of neutralizing PAD4 activity is maintained. Such substitutions, deletions, or additions may be introduced into the CDR, but it is preferable that they be introduced into regions other than the CDR. Furthermore, it is preferable that such amino acid substitutions are conservative substitutions in order to maintain the properties of this disclosure. The amino acid sequence of the anti-PAD4 antibody containing substitutions, deletions, etc. in the heavy chain variable region and / or light chain variable region is such that the heavy chain variable region comprises 90% or more (more preferably 95%) of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 35, or SEQ ID NO: 43. The amino acid sequence may have 90% or more identity (more preferably 95%, 96%, 97%, 98%, 99% or more), and the light chain variable region may have 90% or more identity (more preferably 95%, 96%, 97%, 98%, 99% or more) with the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17, the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19, the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21, the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 23, the amino acid sequence of SEQ ID NO: 28, the amino acid sequence of SEQ ID NO: 36, or the amino acid sequence of SEQ ID NO: 44.An anti-PAD4 antibody may also be an anti-PAD4 antibody in which the heavy chain has an amino acid sequence that is 90% or more (more preferably 95%, 96%, 97%, 98%, 99% or more) identical to sequence numbers 16, 18, 20, 22, 27, 35, 43 in the sequence listing, and the light chain has an amino acid sequence that is 90% or more (more preferably 95%, 96%, 97%, 98%, 99% or more) identical to sequence numbers 17, 19, 21, 23, 28, 36, 44 in the sequence listing.

[0052] In this disclosure, PAD inhibitors such as PAD4 antibodies, anti-PAD2 / PAD4 bispecific antibodies, or antibody fragments thereof may be used alone, or they may be mixed with pharmacologically acceptable carriers to form a composition. The composition may be prepared by any method known in the art of pharmaceuticals. Examples of pharmacologically acceptable carriers include excipients, lubricants, binders, disintegrants, solvents, solubilizers, suspending agents, isotonic agents, buffers, and analgesics. Furthermore, as needed, appropriate amounts of additives such as common preservatives, antioxidants, colorants, sweeteners, adsorbents, and wetting agents may be used as appropriate.

[0053] The administration form of PAD inhibitors or compositions containing PAD inhibitors is not particularly limited and can be administered to mammals, including humans, by any of the following routes of administration: oral administration, parenteral administration (e.g., intravenous injection, intramuscular injection, subcutaneous administration, rectal administration, transdermal administration, intracerebral administration, intrathecal administration, or other local administration).

[0054] Dosage forms for oral and parenteral administration and methods for preparing them are well known to those skilled in the art, and pharmaceutical compositions can be manufactured by combining PAD inhibitors with pharmaceutically acceptable carriers, etc.

[0055] Dosage forms for parenteral administration include injectable preparations (e.g., intravenous infusion, intramuscular injection, subcutaneous injection, intradermal injection, intracerebral administration preparation, intraspinal administration preparation), topical preparations (e.g., ointments, poultices, lotions), suppositories, eye preparations, eye ointments, nasal drops, ear drops, liposomal preparations, etc. In particular, when direct action on the central nervous system is desired, continuous infusion can be performed using a medical micropump with osmotic pressure, or the preparation can be mixed with fibrin glue or the like to create a sustained-release formulation which can then be implanted in the affected tissue. For example, injectable preparations are usually prepared by dissolving antibodies in distilled water for injection, but solubilizers, buffers, pH adjusters, isotonic agents, analgesics, preservatives, stabilizers, etc. can be added as needed. Freeze-dried preparations for immediate preparation are also available. Dosage forms for oral administration include solid or liquid forms, specifically tablets, coated tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, injections, lozenges, and the like.

[0056] The pharmaceutical composition may further contain other therapeutically effective drugs, and may also contain ingredients such as disinfectants, anti-inflammatory agents, vitamins, and amino acids as needed.

[0057] The dosage of PAD inhibitors is determined by a physician based on various factors, such as the route of administration, the type of disease, the severity of symptoms, the patient's age, sex, weight, the severity of the disease, pharmacological knowledge including pharmacokinetic and toxicological characteristics, whether a drug delivery system is used, and whether it is administered as part of a combination of other drugs. Typically, for an adult (60 kg body weight), the oral dose is 1 to 5000 μg / day, preferably 10 to 2000 μg / day, more preferably 50 to 2000 μg / day, and the injectable dose is 1 to 5000 μg / day, preferably 5 to 2000 μg / day, more preferably 50 to 2000 μg / day, administered in one or several divided doses. For systemic parenteral administration, the dose can be 10 to 100,000 μg / kg per body weight, more preferably 100 to 50,000 μg / kg, and even more preferably 500 to 20,000 μg / kg, administered once a day, once a week, once a month, or once to seven times a year. For local administration using an osmotic pump or the like, it can usually be continuously infused at a rate of 10 to 100,000 μg / day, more preferably 100 to 10,000 μg / day, and even more preferably 500 to 5,000 μg / day per adult (body weight 60 kg).

[0058] In this disclosure, the therapeutic, preventive, and / or relapse-preventive effects of PAD inhibitors in rheumatoid arthritis (RA) may be evaluated, for example, by arthritis score, RA score, swelling width, imaging studies, modified Total Sharp score, Disease Activity Score (DAS), ACR20, ACR50, ACR70 (representing the achievement rate of the rheumatoid activity assessment criteria developed by the American College of Rheumatology (ACR)), or disease markers. When evaluating by arthritis score, for example, a therapeutic, preventive, and / or relapse-preventive effect may be determined if the arthritis score of a patient receiving a PAD inhibitor is significantly lower than the arthritis score of a patient not receiving the PAD inhibitor. Alternatively, a therapeutic, preventive, and / or relapse-preventive effect may be determined if the arthritis score of a patient receiving a PAD inhibitor is significantly lower than the arthritis score of a patient receiving a negative control. The above reduction may be, for example, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 point or less, and may be within the range of any two of these values. Alternatively, if the graph area of ​​the arthritis score of a patient administered with a PAD inhibitor is significantly reduced compared to the graph area of ​​the arthritis score when not administered, it may be determined that there was a therapeutic, preventive, and / or relapse-preventive effect. Alternatively, if the graph area of ​​the arthritis score of a patient administered with the composition is significantly reduced compared to the graph area of ​​the arthritis score of a patient administered with a negative control substance, it may be determined that there was a therapeutic, preventive, and / or relapse-preventive effect. The above reduction may be, for example, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or less, or within the range of any two of these values.

[0059] <2. Embodiments in which a PAD inhibitor is administered to subjects whose levels of neutrophil activation-related factors are higher than normal> Another embodiment of the present disclosure is a pharmaceutical for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient, and administered to subjects whose levels of neutrophil activation-related factors in the blood are higher than normal. The autoimmune arthritis is, for example, rheumatoid arthritis. This embodiment can be rephrased as follows: A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor as an active ingredient, and administered to subjects whose levels of neutrophil activation-related factors in the blood are higher than normal. A method for the treatment or prevention of autoimmune arthritis, comprising administering an effective amount of a PAD inhibitor to subjects whose levels of neutrophil activation-related factors in the blood are higher than normal. The use of a PAD inhibitor in the manufacture of a medicament for the treatment or prevention of autoimmune arthritis, wherein the medicament is administered to a subject whose blood level of neutrophil activation-related factor is higher than a reference value; a PAD inhibitor for use in the treatment or prevention of autoimmune arthritis, wherein the PAD inhibitor is administered to a subject whose blood level of neutrophil activation-related factor is higher than a reference value; a method for the treatment, prevention or prevention of relapse of autoimmune arthritis, comprising: measuring the level of neutrophil activation-related factor in the blood of a subject, or receiving the result of measuring the level of neutrophil activation-related factor in the blood of a subject; comparing the level of neutrophil activation-related factor with a reference value; selecting a subject if the level of neutrophil activation-related factor is higher than a reference value; and administering an effective amount of the PAD inhibitor to the selected subject.

[0060] In this disclosure, neutrophil activation-related factors are selected from the group consisting of, for example, PAD protein, myeloperoxidase (MPO) protein, CXCL13 protein, citrullinated protein, and ACPA. PAD may be PAD4.

[0061] In this disclosure, reference values ​​are, for example, levels of neutrophil activation-related factors in the blood of healthy individuals or patients with low disease activity of autoimmune arthritis. That is, reference values ​​can be experimentally determined as thresholds that can distinguish between groups by measuring, for example, the levels of neutrophil activation-related factors in the blood of patients with disease activity of autoimmune arthritis and the levels of neutrophil activation-related factors in the blood of healthy individuals or patients with low disease activity of autoimmune arthritis. The method for determining reference values ​​for the concentrations and activities of PAD protein, MPO protein, CXCL13 protein, citrullinated protein, and ACPA in the blood in this disclosure is not particularly limited and can be determined, for example, using general statistical methods.

[0062] Specifically, as a method for determining reference values, for example, in patients diagnosed with autoimmune arthritis with the desired disease activity using commonly used diagnostic methods for autoimmune arthritis, the level of neutrophil activation-related factors in blood collected before or after diagnosis, such as upon admission, is measured or the measurement results are received. After measuring or obtaining measurement results for multiple patients, the level of neutrophil activation-related factors in the blood of these patients can be calculated from the mean or median, and the value that does not include this factor can be used as the reference value.

[0063] Furthermore, for patients with autoimmune arthritis exhibiting multiple disease activities and for multiple healthy individuals or patients with autoimmune arthritis exhibiting low disease activity, the levels of neutrophil activation-related factors in the blood collected before or after diagnosis, such as at the time of hospitalization, can be measured. The levels of neutrophil activation-related factors in the blood of patients with autoimmune arthritis exhibiting disease activity and the levels of neutrophil activation-related factors in the blood of healthy individuals or patients with autoimmune arthritis exhibiting low disease activity, along with their variability, can be calculated from the mean or median values. Then, a threshold can be determined that distinguishes between the two values, taking variability into consideration, and this threshold can be used as the reference value.

[0064] In this context, "level of neutrophil activation-related factors in the blood" refers to the amount, concentration, and activity (enzyme activity, etc.) of neutrophil activation-related factors in the blood. The level of each factor can be measured using a method appropriate to its type, and may be expressed as, for example, the amount of moles, mass, or number of units of the factor present per 1 mL of blood, or the amount of moles or mass relative to 1 mg of total protein in the blood.

[0065] The measurement methods for each factor are not particularly limited and can be measured using various methods commonly used to quantitatively or semi-quantitatively measure protein expression. Specific measurement methods include, for example, methods utilizing immunoreactions such as radioimmunoassay, enzyme immunoassay, fluorescence immunoassay, luminescence immunoassay, immunoprecipitation, immunoturbidimetry, Western blotting, immunostaining, and immunodiffusion. As for methods for measuring PAD activity, for example, a method of quantifying citrullinated compounds produced by adding a low-molecular-weight substrate using liquid chromatography-mass spectrometry is one example.

[0066] The aforementioned measurement or receipt of measurement results is performed, for example, before or after administering an autoimmune arthritis treatment drug to the subject.

[0067] Pharmaceuticals containing PAD inhibitors may have a statement in their package insert indicating that they are "administered to subjects whose levels of neutrophil activation-related factors in the blood are higher than the normal range."

[0068] The PAD inhibitor and the pharmaceutical or composition containing the same are as described in the first embodiment above.

[0069] <3. Embodiments relating to a method for predicting the effectiveness of treatment or prevention by PAD inhibitors> Another embodiment of the present disclosure is a method for predicting the effectiveness of a composition for the treatment, prevention, or prevention of relapse of autoimmune arthritis, comprising a PAD inhibitor, the method comprising: measuring or receiving the level of neutrophil activation-related factor in the blood of a subject before administration of the composition; measuring or receiving the level of neutrophil activation-related factor in the blood of a subject after administration of the composition; comparing the level of neutrophil activation-related factor measured before administration of the composition with the level of neutrophil activation-related factor measured after administration of the composition; and predicting that the composition is effective in treating, preventing, or preventing relapse of the subject if the level of neutrophil activation-related factor measured after administration of the composition is lower than the level of neutrophil activation-related factor measured before administration of the composition.

[0070] A further embodiment of the present disclosure is a method for predicting the effectiveness of a composition for the treatment, prevention, or prevention of relapse of autoimmune arthritis, comprising a PAD inhibitor, the method comprising: measuring or receiving the level of neutrophil activation-related factor in the blood of a subject before administration of the composition; administering the composition to the subject; measuring or receiving the level of neutrophil activation-related factor in the blood of the subject after administration of the composition; comparing the level of neutrophil activation-related factor measured before administration of the composition with the level of neutrophil activation-related factor measured after administration of the composition; and predicting that the composition is effective in treating, preventing, or preventing relapse of the subject if the level of neutrophil activation-related factor measured after administration of the composition is lower than the level of neutrophil activation-related factor measured before administration of the composition.

[0071] The neutrophil activation-related factors are selected from the group consisting of, for example, activated PAD, activated PAD4, PAD4 protein, MPO protein, CXCL13 protein, citrullinated protein, and ACPA. The measurement of neutrophil activation-related factors is as described above. The autoimmune arthritis is, for example, rheumatoid arthritis. The PAD inhibitor is as described above, but can be, for example, an anti-PAD antibody or an antibody fragment thereof, or an anti-PAD4 antibody or an antibody fragment thereof.

[0072] The period after administration of the composition can be one week, two weeks, three weeks, or four weeks after administration (or the final administration in the case of multiple administrations). If the level of neutrophil activation-related factors measured after administration of the composition is lower than the level of neutrophil activation-related factors measured before administration of the composition, it can be predicted that the composition containing the PAD inhibitor will be effective in treating or preventing autoimmune arthritis. Therefore, the composition containing the PAD inhibitor can be administered to patients suffering from autoimmune arthritis or to subjects at high risk of developing autoimmune arthritis.

[0073] <4. Embodiments relating to pharmaceuticals containing PAD inhibitors and TNF inhibitors> Another embodiment of the present disclosure is a pharmaceutical for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor and a TNF inhibitor. This embodiment can be rephrased as follows: A composition for the treatment or prevention of autoimmune arthritis, comprising a PAD inhibitor and a TNF inhibitor as active ingredients. A method for the treatment or prevention of autoimmune arthritis, comprising administering an effective amount of a PAD inhibitor and a TNF inhibitor to a subject in need thereof. Use of a PAD inhibitor and a TNF inhibitor in the manufacture of a pharmaceutical for the treatment or prevention of autoimmune arthritis. A combination of a PAD inhibitor and a TNF inhibitor for use in the treatment or prevention of autoimmune arthritis.

[0074] In this embodiment, the autoimmune arthritis is, for example, rheumatoid arthritis. The PAD inhibitor can be an anti-PAD antibody or an antibody fragment thereof, as described above, for example, an anti-PAD4 antibody or an antibody fragment thereof can be used. Specific examples are also as described above, and an anti-PAD4 antibody containing a specific heavy chain CDR sequence and a light chain CDR sequence, or an anti-PAD4 antibody containing a specific heavy chain variable region sequence and a light chain variable region sequence, or an anti-PAD4 antibody containing a heavy chain sequence and a light chain sequence can be used.

[0075] TNF inhibitors suppress the effects of TNF in vivo and are either anti-TNF antibodies or tumor necrosis factor-alpha / lymphotoxin-alpha (TNFα / LTα) receptor-Fc fusion proteins, more specifically including infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab. Anti-TNF antibodies are preferably anti-TNF-α antibodies.

[0076] In this embodiment, the PAD inhibitor and the TNF inhibitor may be administered separately, or they may be administered as a pharmaceutical composition formulated by pre-mixing the PAD inhibitor and the TNF inhibitor.

[0077] In an embodiment in which the PAD inhibitor and the TNF inhibitor are administered separately, the dose of the PAD inhibitor is preferably 1 to 5000 μg / day in the case of an anti-PAD antibody, and the dose of the TNF inhibitor is preferably 1 to 5000 μg / day in the case of an anti-TNF antibody.

[0078] The timing of administration of PAD inhibitors and TNF inhibitors does not matter; they may be administered first, followed by TNF inhibitors, followed by PAD inhibitors, or simultaneously. However, it is preferable to administer TNF inhibitors first, followed by PAD inhibitors. PAD inhibitors and TNF inhibitors may be administered alternately. PAD inhibitors and TNF inhibitors may be pre-packaged in kits, and the respective dosages and timings of administration may be described in the package insert.

[0079] In a pharmaceutical composition containing a PAD inhibitor and a TNF inhibitor, for example, one dose of the preparation should contain an amount such that the single dose of the PAD inhibitor is 1 to 5000 μg in the case of an anti-PAD antibody, and an amount such that the single dose of the TNF inhibitor is 1 to 5000 μg in the case of an anti-TNF antibody. As described in the first embodiment above, such a pharmaceutical composition can be mixed with a pharmacologically acceptable carrier as needed and formulated into an appropriate dosage form such as an injection for use.

[0080] The invention will be described in detail below with reference to examples, but this disclosure is not limited to the contents of the examples described below.

[0081] Example 1 Plasma concentrations of PAD4 protein and ACPA in rheumatoid arthritis patients Rheumatoid arthritis patients with a Disease Activity Score-28 for Rheumatoid Arthritis with Erythrocyte Sedimentation Rate (DAS28-ESR) of 3.2 or higher before the initiation of biological antirheumatic drug treatment were evaluated. Based on their treatment history, they were classified into three groups: patients who responded to TNF inhibitors (TNF-R, 6 patients), patients who did not respond to TNF inhibitors (TNF-IR, 8 patients), and patients who did not respond to biological antirheumatic drugs other than TNF inhibitors (non-TNF-IR, 8 patients). TNF inhibitors are any of the following drugs: Adalimumab, Certolizumab, Pegol, Etanercept, Golimumab, or Infliximab. Other biological antirheumatic drugs are Tocilizumab or Abatacept. Patients with a DAS28-ESR score below 2.5 for at least three months of treatment were classified as responders, and patients with a DAS28-ESR score of 3.2 or higher were classified as non-responders. Most patients had been prescribed methotrexate before treatment with biological antirheumatic drugs.

[0082] For each patient sample, plasma PAD4 protein concentration and plasma anti-citrullinated peptide antibody (ACPA) concentration were quantified using commercially available ELISA kits before (pre) and after (Post) treatment with biological anti-rheumatic drugs (using the most recent sample collected over time during the treatment period, not necessarily indicating completion of treatment). The PAD4 (human) ELISA Kit (Cayman Chemical, 501460) was used to measure PAD4 protein concentration, and the ACPA ELISA Kit (Abnova, KA1268) was used to measure ACPA concentration.

[0083] The results are shown in Figure 1. PAD4 protein concentrations were higher in TNF-IR and non-TNF-IR patients compared to healthy individuals (HC) and TNF-R patients (especially pre-TNF-R). ACPA concentrations were also higher in TNF-IR and non-TNF-IR patients compared to healthy individuals (HC) and TNF-R patients.

[0084] Example 2 Positive correlation between the amount of neutrophil activation-related factors and DAS28-ESR In each patient from Example 1, plasma obtained over time was used to quantify plasma myeloperoxidase (MPO) protein concentration, plasma PAD4 protein concentration, and plasma anti-citrullinated peptide antibody (ACPA) concentration using commercially available ELISA kits. For measuring MPO protein concentration, the Human MPO ELISA Kit (Proteintech, KE00171) was used; for measuring PAD4 protein concentration, the PAD4 (human) ELISA Kit (Cayman Chemical, 501460) was used; and for measuring ACPA concentration, the ACPA ELISA Kit (Abnova, KA1268) was used. In addition, plasma PAD activity was measured by quantifying citrullinated compounds generated by adding a low-molecular-weight substrate to plasma using liquid chromatography-mass spectrometry. For more detailed information on PAD activity measurement, 80 μL of PAD assay buffer (50 mM HEPES, 10 mM CaCl2, pH 7.7) was added to each 96-well plate, followed by 2.5 μL of plasma sample. After incubation at room temperature for 5 minutes, 5 μL of benzoyl-arginine ethyl ester (BAEE, Cayman Chemical, 700563) was added as a substrate for PAD4. After incubation at 37 °C for 24 hours, 10 μL was transferred to a 96-well plate, 100 μL of 80% acetonitrile solution was added, and the precipitate was removed by centrifugation. The concentration of the product (benzoyl-citrulline ethyl ester, BCEE) in the supernatant was quantified using a QTRAP6500 LC-MS / MS system. Pearson's correlation coefficient between MPO protein concentration, PAD4 protein concentration, ACPA concentration, or PAD activity and DAS28-ESR was calculated at each time point. Compared to patients with TNF-R, patients with TNF-IR and non-TNF-IR showed a tendency towards a positive correlation between DAS28-ESR and any of the following: MPO protein concentration, PAD4 protein concentration, ACPA concentration, or PAD activity.The correlation coefficient between ACPA concentration and DAS28-ESR in TNF-IR was 0.483, the correlation coefficient between PAD activity and DAS28-ESR in non-TNF-IR was 0.708, and the correlation coefficient between PAD4 protein concentration and DAS28-ESR in non-TNF-IR was 0.406, indicating significant positive correlations.

[0085] Example 3: Action of anti-PAD4 antibody in a collagen-induced arthritis (CIA) model. CIA model mice were prepared by immunizing 6-8 week old female DBA / 1 mice with bovine type II collagen and Freund's adjuvant twice: on day 1 and 21 days after the initial immunization. The antibody described in International Publication No. 2022 / 176970 was used as the anti-PAD4 antibody. The sequences of the variable regions of the heavy and light chains of the antibody used were the amino acid sequences from amino acid numbers 1-120 of SEQ ID NO: 20 and amino acid sequences from amino acid numbers 1-105 of SEQ ID NO: 21, respectively. The heavy and light chain sequences are SEQ ID NOs: 20 and 21. 29 days after the initial immunization, mice that developed arthritis were administered anti-PAD4 antibody intravein at doses of 15 mg / kg, 6 mg / kg, or 3 mg / kg (8-9 mice in each group). The same doses of anti-PAD4 antibody were administered again 36 and 43 days after the initial immunization. Etanercept was administered intraperitoneally every two days at a dose of 10 mg / kg 29 days after the initial immunization (8 mice in each group). The arthritis score was evaluated according to (i) to (iii) below: (i) The evaluation sites were each finger, back of the hand, and joint of the limb. (ii) The arthritis score was assigned according to Table 1. (iii) The arthritis score was expressed as the sum of the scores for each finger, back of the hand, and joint of the limb (maximum value was 16 / mouse).

[0086] Homogenates were prepared from the limbs 50 days after the initial immunization, and the amount of citrullinated protein, i.e., the amount of citrulline per unit of protein, was quantified. The quantification was performed using the following procedure: 100% trichloroacetic acid (TCA) aqueous solution was added to the homogenate in a 4:1 ratio, and after cooling on ice, the mixture was centrifuged to separate the protein pellet. Citrulline quantification was performed according to the Homocitrulline / Citrulline Assay Kit (Cell Biolabs, MET-5027) protocol. 50 μL of PBS was added to the TCA precipitate, and 5 μL each of SDS Solution and roteinase K Solution were added, and the mixture was reacted at 37°C for 2 hours. 250 μL of Assay Reagent A and 50 μL of Assay Reagent B were added, and the mixture was heated at 95°C for 30 minutes. The insoluble fraction was centrifuged, and the absorbance of the supernatant at 540 nm was measured using a microplate spectrophotometer (SpectraMax M2e, Molecular Devices). On the same day, serum concentrations of CXCL13 protein and ACPA were quantified using commercially available ELISA kits. For CXCL13 protein concentration, the Mouse CXCL13 / BLC / BCA-1 Quantikine ELISA Kit (R&D Systems, MCX130) was used. For ACPA concentration, the MESACUP-2 test plate (Laboratory Institute of Medicine and Biology) and HRP-labeled anti-mouse IgG antibody for detection were used.

[0087] The results are shown in Figure 2. Anti-PAD4 antibodies significantly reduced arthritis score and citrullinated protein levels, CXCL13 protein concentration, and ACPA concentration at all doses (arthritis score was evaluated by Shirley-Williams multiple comparison test. Citrullinated protein levels, CXCL13 protein concentration, and ACPA concentration were evaluated by Williams multiple comparison test. *p<0.025, **p<0.005). Etanercept suppressed arthritis score to a similar extent as anti-PAD4 antibodies, but did not suppress citrullinated protein levels, CXCL13 protein concentration, or ACPA concentration (arthritis score was evaluated by Wilcoxon test. Citrullinated protein levels, CXCL13 protein concentration, and ACPA concentration were evaluated by Student's t-test. *p<0.05, **p<0.01).

[0088]

[0089] Example 4: Effect of anti-PAD4 antibody in Etanercept-unresponsive CIA model mice A CIA model was created using 5-7 week old female DBA / 1 mice in the same manner as in Example 3. Twenty-eight days after the initial immunization, all mice with an arthritis score of 0 were administered etanercept intraperitoneally at a dose of 10 mg / kg every two days. At 34, 36, and 38 days after the initial immunization, etanercept-unresponsive mice that developed arthritis with an arthritis score of 1 or higher after administration of etanercept were divided into two groups (11 mice in each group). In one group, etanercept was administered every two days for seven doses until two days before the end of the study. In the other group, etanercept was not administered thereafter, and instead, anti-PAD4 antibody was administered intratail vein at a dose of 0.3 mg / kg on the day of group division. The same dose of anti-PAD4 antibody was also administered one week after the initial administration. The anti-PAD4 antibody used was the anti-PAD4 antibody described in Example 3. For mice that received continuous administration of Etanercept and mice that received the anti-PAD4 antibody, the arthritis score described in Example 3 was measured every two days for 14 days when administration of Etanercept was replaced with the anti-PAD4 antibody, and the average score for all time points was calculated.

[0090] The results are shown in Figure 3. Administration of anti-PAD4 antibody to etanercept-unresponsive mice resulted in a reduction in arthritis scores.

[0091] Example 5: Combined effect of Etanercept and anti-PAD4 antibody in CIA model mice. A CIA model was created using 5-7 week old female DBA / 1 mice in the same manner as in Example 3. 28 days after initial immunization, mice that developed arthritis were administered either Etanercept alone, anti-PAD4 antibody alone, or both Etanercept and anti-PAD4 antibody. Etanercept was administered intraperitoneally every two days at a dose of 10 mg / kg (12 mice in each group). Anti-PAD4 antibody was administered intratail vein at a dose of 0.3 mg / kg 28, 35, and 42 days after initial immunization (12 mice in each group). The anti-PAD4 antibody used was the one described in Example 3. The arthritis score described in Example 3 was measured every two days.

[0092] The results are shown in Figure 4. Combining etanercept with an anti-PAD4 antibody significantly reduced arthritis scores compared to administering each drug alone (arthritis scores were evaluated at each time point using Student's t-test; *p<0.05, **p<0.01).

[0093] Example 6: PAD4 Immunostaining in CIA Model Mouse Joints A CIA model was prepared in the same manner as in Example 3 using 6-8 week old female DBA / 1 mice. Hind limbs were collected 28, 35, and 42 days after the initial immunization, fixed with formalin, and paraffin-embedded sections were prepared. The deparaffinized tissue sections were immersed in Target Retrieval Solution, pH 9, pressurized and heated, and then thoroughly air-cooled. After blocking nonspecific protein adsorption, the sections were reacted overnight at 4°C with a diluted solution of Anti-PADI4 / PAD4 antibody [clone:214810] (abcam). After washing the sections, staining was performed using Histofine Simple Stain Mouse MAX-PO(R) (Nichirei) and Peroxidase Staining DAB Kit (Nacalai Tesque).

[0094] The results are shown in Figure 5. Pannus formed in the joint cavity 35 and 42 days after the initial immunization was strongly stained with anti-PAD4 antibody.

[0095] Example 7 Calculation of PAD activity in human plasma Plasma samples from three rheumatoid arthritis patients were each divided into two portions. One portion was left untreated, and the other portion was treated with a large excess of anti-PAD4 antibody. The anti-PAD4 antibody used was the one described in Example 3. Next, the PAD activity value was calculated for all plasma samples using the PAD activity measurement method described in Example 2. As a result, the calculated PAD activity values ​​with and without anti-PAD4 antibody were 1.156 (without) and 0.892 (with), 1.303 (without) and 1.233 (with), and 0.938 (without) and 0.786 (with). The difference between the PAD activity value of the sample without the anti-PAD4 antibody and the PAD activity value of the sample with the anti-PAD4 antibody added was taken as the PAD4 activity value. In this way, PAD4 activity could be calculated.

[0096] In light of the above teachings, it is clear that the present disclosure is subject to numerous modifications and variations. Therefore, it should be understood that, within the scope of the appended claims, the present disclosure may be implemented in ways different from those specifically described herein.

Claims

1. A pharmaceutical product for the treatment or prevention of autoimmune arthritis, comprising a peptidylarginine deiminase (PAD) inhibitor as an active ingredient, and administered to subjects unresponsive to or expected to be unresponsive to at least one disease-modifying antirheumatic drug.

2. The pharmaceutical product according to claim 1, wherein the subjects predicted to be unresponsive to the disease-modifying antirheumatic drug are subjects whose levels of neutrophil activation-related factors in the blood are higher than the reference value.

3. The pharmaceutical product according to claim 2, wherein the reference value is the level of neutrophil activation-related factor in the blood of a healthy person or a patient with low disease activity of autoimmune arthritis.

4. The pharmaceutical product according to claim 1, wherein the disease-modifying antirheumatic drug is a synthetic antirheumatic drug or a biological antirheumatic drug.

5. The pharmaceutical product according to claim 4, wherein the synthetic antirheumatic drug is a conventional synthetic antirheumatic drug or a molecularly targeted synthetic antirheumatic drug.

6. The pharmaceutical product according to claim 5, wherein the conventional synthetic antirheumatic drug is methotrexate.

7. The pharmaceutical product according to claim 5, wherein the molecularly targeted synthetic antirheumatic drug is a Janus kinase inhibitor.

8. The pharmaceutical product according to claim 4, wherein the biological antirheumatic drug is a tumor necrosis factor (TNF) inhibitor, an interleukin (IL)-6 inhibitor, or a T cell selective costimulation modulator.

9. The pharmaceutical product according to claim 8, wherein the TNF inhibitor is an anti-TNF antibody or a TNF-alpha / lymphotoxin-alpha receptor-Fc fusion protein.

10. The pharmaceutical product according to claim 8, wherein the IL-6 inhibitor is an anti-IL-6 receptor antibody.

11. The pharmaceutical product according to claim 8, wherein the T cell selective costimulation modifier is a cytotoxic T-lymphocyte antigen-4-Fc fusion protein.

12. The pharmaceutical product according to claim 1, wherein the autoimmune arthritis is rheumatoid arthritis.

13. The pharmaceutical product according to any one of claims 1 to 12, wherein the PAD inhibitor is an anti-PAD antibody or a fragment of that antibody.

14. The anti-PAD antibody or antibody fragment thereof is: a-1) an anti-PAD4 antibody or antibody fragment thereof in which the complementarity determining region (CDR) 1 of the heavy chain contains the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain contains the amino acid sequence of SEQ ID NO: 7, the CDR3 of the heavy chain contains the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain contains the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain contains the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain contains the amino acid sequence of SEQ ID NO: 13; b-1) an anti-PAD4 antibody or antibody fragment thereof in which the CDR1 of the heavy chain contains the amino acid sequence of SEQ ID NO: 1, the CDR2 of the heavy chain contains the amino acid sequence of SEQ ID NO: 8, the CDR3 of the heavy chain contains the amino acid sequence of SEQ ID NO: 11, the CDR1 of the light chain contains the amino acid sequence of SEQ ID NO: 4, the CDR2 of the light chain contains the amino acid sequence of SEQ ID NO: 5, and the CDR3 of the light chain contains the amino acid sequence of SEQ ID NO:

13. c-1) an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 14; and d-1) an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:

15.

15. The anti-PAD antibody or antibody fragment thereof is: a-2) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 13; b-2) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 8, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 13; c-2) An anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 14; and d-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:

15.

16. The anti-PAD antibody or antibody fragment thereof is: a-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO:

23. A pharmaceutical product according to claim 13, selected from the group consisting of the following.

17. The anti-PAD antibody or antibody fragment thereof is: a-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-4) A pharmaceutical product according to claim 13, selected from the group consisting of an anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22, and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO:

23.

18. A pharmaceutical product for the treatment or prevention of autoimmune arthritis, comprising a peptidylarginine deiminase (PAD) inhibitor and a tumor necrosis factor (TNF) inhibitor.

19. The pharmaceutical product according to claim 18, wherein the autoimmune arthritis is rheumatoid arthritis.

20. The pharmaceutical product according to claim 18, wherein the PAD inhibitor is an anti-PAD antibody or a fragment of that antibody.

21. The anti-PAD antibody or antibody fragment thereof is: a-1) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 contains the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 contains the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR3 contains the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 contains the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 contains the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 contains the amino acid sequence of SEQ ID NO: 13; b-1) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 contains the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 contains the amino acid sequence of SEQ ID NO: 8, the heavy chain CDR3 contains the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 contains the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 contains the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 contains the amino acid sequence of SEQ ID NO:

13. c-1) an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 14; and d-1) an anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:

15.

22. The anti-PAD antibody or antibody fragment thereof is: a-2) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 13; b-2) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 8, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 13; c-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 9, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 12, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO: 14; and d-2) An anti-PAD4 antibody or antibody fragment thereof, selected from the group consisting of: an anti-PAD4 antibody or antibody fragment thereof, wherein the heavy chain CDR1 consists of the amino acid sequence of SEQ ID NO: 1, the heavy chain CDR2 consists of the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR3 consists of the amino acid sequence of SEQ ID NO: 11, the light chain CDR1 consists of the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 consists of the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 consists of the amino acid sequence of SEQ ID NO:

15.

23. The anti-PAD antibody or antibody fragment thereof is: a-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-3) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region contains the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22 and the light chain variable region contains the amino acid sequence of amino acids 1 to 105 of SEQ ID NO:

23. A pharmaceutical product according to claim 20, selected from the group consisting of the following.

24. The anti-PAD antibody or antibody fragment thereof is: a-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 16 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 17; b-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 18 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 19; c-4) an anti-PAD4 antibody or antibody fragment thereof in which the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 20 and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO: 21; and d-4) A pharmaceutical product according to claim 20, selected from the group consisting of an anti-PAD4 antibody or an antibody fragment thereof, wherein the heavy chain variable region consists of the amino acid sequence of amino acids 1 to 120 of SEQ ID NO: 22, and the light chain variable region consists of the amino acid sequence of amino acids 1 to 105 of SEQ ID NO:

23.

25. The pharmaceutical product according to any one of claims 18 to 24, wherein the TNF inhibitor is a composition comprising an anti-TNF antibody or a TNF-alpha / lymphotoxin-alpha receptor-Fc fusion protein.

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