Use of SE-DR affinity peptides for the preparation of drugs to treat rheumatic diseases

Abstract

This invention provides the use of SE-DR (HLA-DR molecule containing a co-epitope) affinity peptides for the preparation of drugs to treat rheumatic diseases in rheumatoid arthritis patients who are tuberculosis-positive and / or have hepatitis B. [Solution] The present invention provides a pharmaceutical composition comprising an SE-DR affinity peptide and a non-antigen-specific antirheumatic drug, and the use thereof for the preparation of drugs for the treatment of various types of rheumatic diseases. The pharmaceutical composition can rapidly inhibit the progression of rheumatic diseases and achieve sustained remission even after tapering or discontinuation of the drug, for complete control of rheumatic diseases.

Description

[Technical Field]

[0001] (Cross-reference of related applications) This application is China Patent Application No. 202110873116, filed on July 30, 2021. We claim the benefits of patent no. 6. Chinese Patent Application No. 202110873116.6 is specified herein. It is incorporated as a reference throughout the whole.

[0002] (Technical field) This invention belongs to the field of biopharmaceuticals, and in particular to patients who are tuberculosis positive and / or have hepatitis B. SE-DR parent in the preparation of drugs to treat rheumatic diseases in rheumatoid arthritis patients This invention relates to the use of harmonious peptides. Furthermore, the present invention relates to SE-DR affinity peptides and non-antigen-specific A pharmaceutical composition containing heterologous antirheumatic drugs, and for treating various types of rheumatic diseases. This also relates to its use in the preparation of eye medications. [Background technology]

[0003] Rheumatic diseases are extremely common clinical conditions characterized by joint pain and fear of wind and cold. It is a group of syndromes. Rheumatism is an abbreviation for rheumatic disease, and generally refers to diseases affecting the bones, joints, and muscles. A major disease affecting the flesh and surrounding soft tissues such as bursae, tendons, fascia, blood vessels, and nerves. It refers to a group. Rheumatoid arthritis (RA) is one of the most common rheumatic diseases. Rheumatism is a chronic systemic autoimmune disease characterized by erosive synovitis, affecting the articular cartilage and It causes bone destruction, leading to joint deformation and ultimately disability. The 3-year disability rate for patients without medical care is high at 75%, and it is one of the major causes of labor force loss among young and middle-aged people. It is said that the average lifespan of patients is 3 to 10 years shorter than that of healthy people, hence it is called "immortal cancer." It is called "[...]. This disease causes serious social and economic problems. Onset Initially, they can still work, but after 10 years, 50% of patients lose their ability to work, and after 15 years... It reaches 67%. Rheumatoid arthritis is known as "5D" overseas, meaning disease, depression, and disability. It is said to cause death and debt. The global prevalence of rheumatoid arthritis is 0. The prevalence is 5-1%, and in China it is about 0.3%, with approximately 5 million patients. The peak age of onset for female patients is 35-44 years old, while for male patients it is 55-64 years old. Yes, and the incidence rate increases with age. It is more common in women, with a female-to-male ratio of 2:1-3. :1.

[0004] The pathogenesis of rheumatic diseases is extremely complex. Under the influence of environmental or genetic factors, Pathogenic antigens in susceptible populations activate the innate immune system, and antigens are transmitted to antigen-presenting cells (APCs). It is presented by and activates T cells. T cell activation involves two independent signals. Co-stimulation by is necessary. The first signal is "human leukocyte antigen (HLA)-antigen peptide This involves the formation of a three-molecule complex of the T-cell receptor (TCR), HLA, and antigen peptides. First, it forms a molecular complex, and then it associates with TCR to form a trimolecular complex. The signaling pathway involves the binding of CD28 on the surface of T cells to CD80 / 86 on the surface of APC cells. Activated T cells can further activate B cells, and these activated T cells Cells and B cells enter the peripheral circulation and contain autoantigens that have a structure similar to pathogenic antigens. It reaches the site of mutation. Since the autoantigen functions as a pathogenic antigen, it possesses T cells and B cells, etc. It is continuously activated, secreting large amounts of inflammatory cytokines, chemokines, and collagenase, and lubrication It promotes the over-formation of the membrane, pannus formation and persistent inflammation, and ultimately causes bone destruction. T lymphocytes play an important role in the onset, persistence and recurrence of rheumatoid arthritis. In particular, the differentiation from activated T cells to helper T cells is playing an increasingly important role in the pathogenesis of rheumatoid arthritis and the research of therapeutic drugs. In this field, the changes in the levels of Th17 cells and Treg cells have become the focus of research.

[0005] Currently, the therapeutic drugs for rheumatic diseases (especially RA) can be divided into four generations according to their development times and mechanisms. The first generation is non-steroidal anti-inflammatory drugs (NSAIDs), the second generation is glucocorticoids (GCs), the third generation is disease-modifying anti-rheumatic drugs (DMARDs), and the fourth generation is biological agents represented by TNF-α inhibitors. The fourth-generation therapeutic drugs for rheumatic diseases include TNF-α inhibitors, IL-1 antagonists, IL-6 antagonists, JAK3 inhibitors, T cell inhibitors, B cell inhibitors, etc., and currently occupy up to 97% of the market in Japan, the United States and Europe. However, the drugs commonly used clinically

[0006] as therapeutic drugs for rheumatic diseases (especially RA) have major drawbacks in terms of safety and efficacy. The median lethal dose of methotrexate, which is the first-choice drug recommended clinically, in rats is 43 mg / kg, and its main toxicities are bone marrow suppression and gastrointestinal toxicity. The preclinical toxicity of etanercept, one of the second-choice drugs clinically, It induces cancer and increases the incidence of cancer. Preclinical safety study on the carcinogenicity of abatacept In the clinical trial, approximately 50% of the mice in the treatment group died from lymphoma, and the incidence of breast cancer also increased. In trials, abatacept has been shown to worsen infections and increase the risk of cancer. Yes, most biological agents (such as TNF inhibitors) carry a risk of tumors and infections. Because the FDA has issued a black box warning, patients should undergo a tuberculin test before administration. Patients are required to undergo testing for hepatitis B and related conditions. Patients who test positive for tuberculin are required to... Even if a biological agent is administered after tuberculosis treatment and the tuberculin test is negative, the administration of biological agents may still be necessary. During treatment, be vigilant for active tuberculosis. For patients with hepatitis B complications, HBcAg-positive patients should be treated with ritodrine. When initiating ximab therapy or targeted synthetic DMARD therapy, prophylactic anticoagulant Viral therapy should be administered. HBcAb-positive / HBcAg-negative patients should not undergo non-rituximab therapy. When initiating biological agents or targeted synthetic DMARD therapy, careful monitoring is necessary. Therefore, for tuberculosis-positive patients or patients with hepatitis B, the drug options are limited. This presents the problem of increased medication risks.

[0007] Furthermore, currently, a two-year treatment program using different regimens and different criteria for common clinical drugs is underway. Subsequent follow-up studies showed that the remission rate worldwide was only about 40%, and nearly half of the patients did not achieve effective results. It is uncontrollable. In China, the clinical remission rate for RA patients varies depending on the clinical criteria after treatment. When followed up, the percentage was only 8.6-25.2%. A considerable number of patients are currently unable to treat their condition. Therapies have not achieved clinical remission or disease control.

[0008] Safety concerns of existing clinical drugs that can alleviate rheumatic diseases (especially RA) The common reason can be attributed to "insufficient specificity to the target." NSAIDs, GCs and DMARDs have broad immunosuppressive effects, but they specifically target disease-related lymphocytes. It cannot inhibit it. Biological agents and novel small molecule drugs are targeted therapies for RA, Most of these studies target cytokines and kinases downstream of the pathogenesis of rheumatoid arthritis (RA). While inhibiting the aforementioned factors, inhibiting TNF-α increases the incidence of tuberculosis infection and tumors. It can cause side effects such as [unclear]. Abatacept activates T cells, which are at the very top of the rheumatoid arthritis (RA) development pathway. It is effective in this regard. Its safety is superior to that of TNF-α inhibitors and DMARDs, The target is not disease-specific T cells, but rather the activation of a broad range of T cells, thus addressing immunodeficiency. This may cause and further increase the incidence of serious infections and cancer during treatment. Therefore, the development of RA treatments that target disease-specific targets solves the safety problems of existing drugs. This is the main route.

[0009] SE-DR affinity peptides (FNS007, etc.) are innovative peptides developed independently in China. It is a highly effective drug that acts directly on T lymphocytes and is the primary treatment for rheumatic diseases (especially RA). It can be considered a fifth-generation drug. SE-DR affinity peptide is a rheumatic disease-related antigen peptide. By competitively inhibiting the binding of butyl to major histocompatibility complex molecules, its role is... Indeed, the first signal of specific T cell activation mediated by rheumatic disease-associated antigens is 3 By inhibiting the formation of the molecular complex "HLA-antigen peptide-T lymphocyte receptor (TCR)" This antigen-specific therapeutic agent can achieve the therapeutic objectives of rheumatic diseases (especially RA). To achieve this, it inhibits the activation and proliferation of disease-specific autoreactive T cells, and promotes T cell differentiation and development. It can alter the secretion of cytokines. "Antigen-specific therapy" is SE-DR parent This is a key characteristic of the mechanism of harmonious peptides. SE-DR affinity peptides are primarily RAn-based. It is a peptide ligand that modifies essential self-antigen epitopes, and is specific to the development and onset of RA. It acts at a critical stage, inhibiting the binding of various RA-related antigenic peptides to HLA molecules. This can control the patient's disrupted immune microenvironment, reducing inflammatory cells and suppressing inflammation cells. By increasing this factor, immune tolerance is restored, and long-term improvement completely reverses the progression of the disease. It is expected to prevent this. SE-DR affinity peptides in the treatment of rheumatic diseases (for example) The mechanism of FNS007 can be said to indicate the future direction of development for anti-rheumatic drugs. .

[0010] FNS007 is a new class developed by Hebei Fitness Biotechnology Co., Ltd. 1.1 This invention is a pharmaceutical product and has entered the clinical research stage, currently in Phase 1 clinical research. Regarding the new applications of SE-DR affinity peptides (especially FNS007) by the company, existing This research focuses on addressing the safety and efficacy shortcomings of rheumatic disease treatments. Therefore, the object of the present invention is tuberculosis-positive and / or rheumatoid arthritis patients complicated with hepatitis B. This provides the use of SE-DR affinity peptides in the preparation of therapeutic drugs for rheumatic diseases. The objective of the present invention is to use SE-DR affinity peptides and non-antigenic anti-rheumatic This invention provides pharmaceutical compositions containing drugs, and their use in the preparation of therapeutic drugs for various rheumatic diseases. That is what it is. [Overview of the Initiative]

[0011] How do existing rheumatic disease treatments strike a balance between clinical safety and efficacy? The important thing is how to take it. Increasing the dosage within a certain range can enhance the effectiveness of the drug and improve the remission rate. This may improve performance, but at the same time, it may also increase the incidence of side effects. More disease-specific drugs as treatments, and associated risks in drug use. This will solve the problem of insufficient remission rates of existing drugs and will benefit rheumatic disease patients in China. This is a major avenue and a new approach to satisfying treatment acceptance.

[0012] Antigen-specific therapy is a novel approach to address the clinical shortcomings of the above-mentioned drugs for rheumatic diseases. This is a novel idea. Existing non-antigen-specific therapeutic agents (such as TNF-α inhibitors) are currently available. While it has shown some effectiveness, the challenge in autoimmune diseases lies in how to restore tolerance to self-antigens. This is key to treatment, and by using more specific targets, it is possible to reduce the toxicity of drugs for infections and other conditions. This also becomes possible. Therefore, combination therapy with antigen-specific or non-antigen-specific therapy is possible for rheumatic diseases. This is becoming a new trend in disease treatment.

[0013] To achieve the objectives of the present invention, the following technical proposals are provided.

[0014] In a first embodiment, the present invention provides the use of SE-DR affinity peptides, and SE-D R-affinity peptides are peptide segments that bind to HLA DR molecules that share an epitope. This refers to ment, and is observed in rheumatoid arthritis patients who are tuberculosis positive and / or have hepatitis B. It is used in the preparation of pharmaceuticals for the treatment of rheumatic diseases. Preferably, covalent epidermal TOPE is a 5-amino acid molecule with QK / RRAA at positions 70-74 of the HLA-DRβ chain. He is the chief. SE-DR affinity peptides are antigen peptides associated with rheumatic diseases and It plays a role in competitively inhibiting the binding of HLA-DR molecules, but does not have broad immunosuppressive effects. Furthermore, there is no risk of inducing infections or tumors.

[0015] Alternatively, in the above use, the SE-DR affinity peptide bound to SE-DR The sequence contains core amino acids corresponding to P1-P9, and the P1 site has a hydrophobic side chain. It is a rare or unnatural amino acid with similar properties to amino acids, and the P4 site is nonpolar or polar. and uncharged, polar and charged amino acids and rare or unnatural amino acids having similar properties. be.

[0016] Preferably, the corresponding part in the core sequence of the SE-DR affinity peptide bound to SE-DR The P1 sites are Tyr(Y), Phe(F), Trp(W), Leu(L), Ile(I ), Met(M), Val(V), or Ala(A) are selected, and the P4 site is M et(M), Ala(A), Val(V), Ile(I), Leu(L), Asp(D) Selected from Glu(E), Gln(Q), Ser(S), or Cit.

[0017] More preferably, the core sequence of the SE-DR affinity peptide bound to SE-DR contains the pair The corresponding P1 site is selected from Tyr(Y), Phe(F), or Trp(W), and the P4 site The position is selected from Met(M), Leu(L), Asp(D), Glu(E), or Cit. It can be done.

[0018] Alternatively, in the above use, the SE-DR affinity peptide bound to SE-DR The corresponding P6 site in the sequence is an amino acid with a short side chain and possesses similar properties. It is a rare or non-natural amino acid, and the P9 site is an amino acid with a moderate to short side chain. These are rare or non-natural amino acids with similar properties.

[0019] Preferably, the corresponding part in the core sequence of the SE-DR affinity peptide bound to SE-DR The P6 site is Ala(A), Gly(G), Ser(S), Thr(T), or Asn( Selected from N), the P9 site is Ala(A), Gly(G), Leu(L), or Met Selected from (M).

[0020] Alternatively, in the above use, the SE-DR affinity peptide contains the following amino acid sequence. Mu: FXGEQGXXGE, or FXGEXAXXGE, where X is P, K, Q, A or Selected from G.

[0021] Preferably, X is selected from A or G. More preferably, SE-DR affinity peptide The code is FNS007(FKGEQAGAGE).

[0022] Alternatively, in the above use, the SE-DR affinity peptide contains the following amino acid sequence. mu:YXKQXTXXLA, where X is selected from V, A, G, N, L, or K.

[0023] Preferably, X is selected from A or G. More preferably, SE-DR affinity peptide The amino acid sequence of is PKYVKQNTLKLAT, PGYVKQGTLGLAT, YV KQNTLKLA, YVAQNTLKLA, YAKQATLKLA, or YAKQATL Selected from the group consisting of ALA.

[0024] Alternatively, in the above use, the SE-DR affinity peptide contains the following amino acid sequence. mu:IWYIXCFXCEXHXXL, where X is A, G, M, T, V, N, Q or S They are selected.

[0025] Preferably, X is selected from A or G. More preferably, SE-DR affinity peptide The amino acid sequences are IWYINCFGCETHAML, IWYIQCFGCETHAM L, IWYISCFGCETHAML, IWYITCFGCETHAML, IWYINC Select either FACETHAML or IWYINCFVCETHAML.

[0026] Alternatively, in the above use, the SE-DR affinity peptide comprises the following amino acid sequence :RSFXLAXSXXGVG, where X is selected from A, G, T, S or E.

[0027] Preferably, X is selected from A or G. More preferably, SE-DR affinity peptide The amino acid sequence of the RS is RSFLASSETGVG, RSFALASSETGVG, RS FTAASSETGVG, RSFTLDSSETGVG, RSFTLAASETGVG, RSFTLASSATGVG, RSFTLASSEAGVG, or RSFTLDSSET Selected from GVG.

[0028] Alternatively, in the above use, the SE-DR affinity peptide comprises the following amino acid sequence :SAVXLCitXSXXGVR, where X is selected from A, G, R, S, V or P ru.

[0029] Preferably, the amino acid sequence of the SE-DR affinity peptide is SAVRLCitSSVP GVR, SAVELCitSSVPGVR, SAVDLCitSSVPGVR, SAVG LCitSSVPGVR, SAVRLCitFSVPGVR, SAVRLCitSSVE GVR, SAVRLCitSSVKGVR, SAVRLCitSSVWGVR, SAVR LCitWSVPGVR, SAVRLCitSSVRGVR, SAVRLCitKSVP It is GVR, SAVALCitSSVPGVR, or SAVRLCitRSVPGVR. .

[0030] Alternatively, the SE-DR affinity peptide contains the following amino acid sequence: GVYXTCi tXSXXCitLCit, where X is selected from A, G, or V. Preferably, X is Selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide is , GVYATCitSSAVCitLCit.

[0031] Alternatively, the SE-DR affinity peptide contains the following amino acid sequence: QDXNCit XNXXKNS, where X is selected from A, G, F, I, K, or L. Preferably, X is selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide. This is QDFTNCitANKLKNS.

[0032] Alternatively, the SE-DR affinity peptide contains the following amino acid sequence: VVLLVAT XGCitXRXXSAYQDK, where X is selected from A, G, E, V, or N. More preferably, X is selected from A or G. The amino acid sequence is VVLLVATEGCitVRVNSAYQDK.

[0033] Alternatively, in the above use, the amino acid sequence of the SE-DR affinity peptide is MGPK GRTVIIEQSWGSPKVTK, MGPKGRTVIIEQSLGSPKVTK, SIDLKDKKYKNIGAKLVQDVANNTNEEA, SIDLKDKKYKN IGAKLVQLVANNTNEA, QYMCitADQAAGGLR, LTQCit GSVLR, WYNCitCHAAN, VETCitDGQVI, or VCitLCit Selected from SSVESTCitGRSCitPAPPPACitGLT

[0034] Alternatively, in the above use, rheumatic diseases include rheumatoid arthritis, psoriasis, and psoriatic arthritis. Ankylosing spondylitis, undifferentiated spondyloarthropathy, systemic lupus erythematosus, systemic sclerosis, or collagen disease Selected from.

[0035] In a second aspect, the present invention relates to SE-DR affinity peptides and non-antigen-specific antirheumatic compounds. A pharmaceutical composition for the treatment of rheumatic diseases, comprising a drug and a pharmaceutically acceptable carrier. This provides a combination of SE-DR affinity peptides and non-antigen-specific anti-rheumatic drugs. The volume ratio is in the range of 50:1 to 1:50, and more preferably the weight ratio is 20:1 to 1: 20, 15:1~1:15, 10:1~1:10, 5:1~1:5, 4:1~1:4, 3 You can choose from a ratio of 1-1-3 or 2-1-1-2, and the specific dosage ratio depends on the selected non-antigen. The type of specific antirheumatic drug may be determined by the clinician. SE-DR affinity A peptide segment refers to a peptide segment that binds to an HLA-DR molecule that shares an epitope. Preferably, the covalent epitope is located at amino acid sequence positions 70-74 of HLA-DR. This refers to the ptide segment, all of which express the QK / RRAA sequence. SE-DR affinity peptide D competitively inhibits the binding of antigen peptides associated with rheumatic diseases to HLA-DR molecules. It plays a role in restoring the immune microenvironment, which has been disrupted and harmed within the body, and in restoring the immune balance. The pharmaceutical composition rapidly slows the progression of rheumatic diseases and maintains its effects even after the gradual reduction or discontinuation of the drug. Achieve complete remission.

[0036] Alternatively, in the above pharmaceutical composition, the SE-DR affinity peptide that is bound to SE-DR The core sequence of the protein contains core amino acids corresponding to P1-P9, and the P1 site has a hydrophobic side chain. It is an amino acid that possesses similar properties, and is a rare or unnatural amino acid.

[0037] Preferably, the corresponding part in the core sequence of the SE-DR affinity peptide bound to SE-DR The P1 sites are Tyr(Y), Phe(F), Trp(W), Leu(L), Ile(I ), selected from Met(M), Val(V), or Ala(A), and the P4 site is nonpolar, Amino acids that are polar and uncharged, polar and negatively charged, and rare or similar properties It is a non-natural amino acid. Preferably, it is an SE-DR affinity peptide that binds to SE-DR. The corresponding P4 sites in the core sequence are Met(M), Ala(A), Val(V), and Il e(I), Leu(L), Asp(D), Glu(E), Gln(Q) or Ser(S) Selected from: Correspondence in the core sequence of the SE-DR affinity peptide that binds to SE-DR. The P6 site contains amino acids with short side chains and rare or unnatural amino acids with similar properties. It is an acid, and the P9 site contains amino acids and similar rare properties with medium to short side chains. Alternatively, it is a non-natural amino acid. Preferably, an SE-DR affinity peptide that binds to SE-DR. The corresponding P6 sites in the core sequence of cydoplasm are Ala(A), Gly(G), Ser(S), Selected from Thr(T) or Asn(N), the P9 site is Ala(A), Gly(G) It is selected from Leu(L) or Met(M).

[0038] Non-antigen-specific antirheumatic drugs include drugs that target cytokines and drugs that target B cells. Drugs targeting T cells, drugs targeting JAK kinase, and conventional disease-modifying inhibitors Medications for rheumatism (DMARDs), nonsteroidal anti-inflammatory drugs (NSAIDs), or glucocorticoids One or more drugs are selected.

[0039] Preferably, the non-antigen-specific antirheumatic drug is a tumor necrosis factor (TNF) inhibitor, IL-6 Inhibitors, IL-1 inhibitors, IL-17 inhibitors, RANKL inhibitors, T cell costimulatory signaling Inhibitors, CD20 inhibitors, CD22 inhibitors, BlyS and APRIL inhibitors, BAFF inhibitors Harmful agents, JAK inhibitors, BTK inhibitors, Syk inhibitors, IRAK4 inhibitors, p38 inhibitors, Alternatively, one or more low-molecular-weight immunosuppressants are selected.

[0040] More preferably, non-antigen-specific anti-rheumatic disease therapeutic agents include adalimumab, tocilizma , anakinra, tofacitinib, abatacept, rituximab, or their biosimilars One or more of the following: Miller or its generic equivalent, methotrexate, or leflunomide. Selected.

[0041] Alternatively, in the above drug composition, the SE-DR affinity peptide has the following amino acid sequence Includes: FXGEQGXXGE or FXGEXAXXGE, where X is K, Q, A or G Selected from. Preferably, X is selected from A or G. More preferably, SE-D The R-affinity peptide is FNS007 (FKGEQAGAGE).

[0042] Alternatively, the amino acid sequence of the SE-DR affinity peptide is MGPKGRTVIIEQS WGSPKVTK, MGPKGRTVIIEQSLGSPKVTK, SIDLKDKKY KNIGAKLVQDVANNTNEEA, SIDLKDKKYKNIGAKLVQLV ANNTNEEA, QYMCitADQAAGGLR, LTQCitGSVLR, WYN CitCHAAN, VETCitDGQVI, or VCitLCitSSVESTCit Selected from GRSCitPAPPPACitGLT.

[0043] Alternatively, in the above pharmaceutical composition, pharmaceutically acceptable carriers include binders and surfactants. It contains a solubilizer, stabilizer, lubricant, wetting agent and / or diluent. The dosage form of the pharmaceutical composition is C Capsules, tablets, pills, liquids, powders, granules, fine granules, film-coated tablets, precipitates Antidepressants, lozenges, sublingual preparations, chewing preparations, buccal preparations, pastes, syrups, suspensions, etc. Lixyl preparations, emulsions, topical preparations, ointments, patches, compresses, transdermal formulations, lotions, These are inhalants, aerosols, injections, or suppositories.

[0044] Alternatively, in the above pharmaceutical composition, the pharmaceutical composition is administered orally, intravenously, or subcutaneously. Intramuscular administration, intracerebral administration, intranasal administration, intrapulmonary administration, intra-arterial administration, intra-articular administration, intradermal administration, Various routes of administration, including intravitreous administration, intramedullary injection, intraperitoneal administration, intrathecal administration, or transdermal administration. Suitable for administration.

[0045] Alternatively, in the above pharmaceutical composition, the rheumatic disease is rheumatoid arthritis, juvenile idiopathic rheumatoid arthritis. Arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, undifferentiated spondyloarthropathy, systemic lupus erythematosus The disease is selected from systemic sclerosis, collagen disease, Crohn's disease, and ulcerative colitis.

[0046] Preferably, patients suffering from rheumatic diseases are tuberculosis-positive or tuberculosis-negative, Patients suffering from rheumatic diseases may or may not have hepatitis B.

[0047] Compared to prior art, the present invention has the following beneficial effects: (1) The antigen-specific drug SE-DR affinity peptide (especially FNS007) They were the first to demonstrate that it does not have a significant effect on the immune systems of healthy cynomolgus monkeys and healthy rats. I saw it. (2) SE-DR affinity peptides (especially FNS007) showed better results compared to adalimumab. Tuberculosis development in rats with latent Mycobacterium nucleosal infection type II collagen-induced arthritis (CIA) We discovered for the first time that it significantly suppresses the process. (3) In type II collagen-induced arthritis model mice (CIA) infected with chronic hepatitis B Furthermore, SE-DR affinity peptides (especially FNS007) are more effective in treating hepatitis B liver disease compared to abatacept. We were the first to discover that it significantly reduces the flame activation rate. (4) In CIA rats, antigen-specific drugs (especially FNS007, APL20) and non-anti- Use in combination with a specific drug (such as adalimumab, abatacept, or methotrexate). Therefore, the progression of the disease can be rapidly delayed, and adalimumab, abatacept, or methamphetamine can be used. It is safer than using trexate or similar drugs alone, and remission is particularly sustained even after gradual tapering or discontinuation of the medication. I discovered this for the first time. (5) Using antigen-specific drugs (FNS007, APL20, etc.) in combination with other non-antigen-specific drugs By doing so, rheumatoid arthritis can be completely controlled. [Brief explanation of the drawing]

[0048] [Figure 1] This graph shows the inflammation score at each time point during administration in the CIA rat model of the experiment shown in Example 9. [Figure 2] This is a graph of inflammation scores at each time point during drug discontinuation in the CIA rat model of the experiment shown in Example 9. [Modes for carrying out the invention]

[0049] The present invention will be further described below with reference to specific examples. The examples provided are merely illustrative and not intended to limit the scope of the present invention. It should be understood that this is not the case.

[0050] If specific technical or conditional details are not provided in the examples, please refer to the relevant literature or product specifications. The technology or conditions described in the specifications shall be followed. Manufacturer of the reagents or equipment used. Unless otherwise specified, all of these reagents or equipment can be purchased through normal channels. This is a new product.

[0051] Unless otherwise specified, the experimental methods in the following examples are all conventional methods. Unless otherwise specified, all test materials used in the examples are commercially available.

[0052] In this invention, the polypeptide used is from Hebei Fitness Biotechnology Ltd. The company synthesized it using a solid-phase synthesis method and purified it using high-performance liquid chromatography. Yes. Analysis of this polypeptide by mass spectrometry revealed that the polypeptide sequence is correct and pure The degree was over 95%.

[0053] Part I: In vitro test results of SE-DR affinity peptides The SE-DR molecule is directly related to the pathogenesis of rheumatoid arthritis (RA). The SE-DR molecule is related to type II collagen. (CII), Heat Shock Protein 60 (Hsp60), Cartilage Glycoprotein (HCG) Presenting RA-related autoantigens such as p-39) in vivo and further activating autoreactive T cells This causes the development of rheumatoid arthritis. An important aspect of the treatment strategy for rheumatoid arthritis is self This involves preventing the activation of self-reactive T cells by self-antigen peptides through competitive inhibition. Currently, the main idea is to modify the self-antigen peptide to inhibit the binding of the self-antigen. The goal is not to activate autoreactive T cells, but to do so, modified The peptide ligand not only has a predetermined affinity for SE-DR, but also activates T cells. It is necessary to demonstrate the suppression of sexualization. Based on the above principles, we have developed a series of modified peptides. We designed and synthesized Gando to improve its affinity for SE-DR and its effect on peripheral blood T cells in RA patients. The activation ability was screened. The specific steps and results are shown in Examples 1 and 2.

[0054] Example 1: Determination of polypeptide affinity Reaction system setup: Constitute a 200 μL reaction system in a 96-well plate, and in each well... To 500 nM HLA-DR1, 25 nM fluorescently labeled polypeptide, and 200 μM The test polypeptide was contained in five series dilutions at a 5-fold ratio. The last well used as the quality control well was... No test polypeptide was placed in the wells. In the quality control wells, free polypeptide cells were found. To measure the gunal, a fluorescently labeled polypeptide that does not contain MHCII molecules was used. The solution had a pH of 7.4. Before use, add the protease inhibitor cocktail and incubate at 37°C for 30 minutes. The incubation period was 1 minute. The degree of fluorescence change was read. The mean value of the free labeled polypeptide was F. P_free was defined as the value of the well that did not contain the competing peptide, and FP_no_comp was defined as the value of the well that did not contain the competing peptide. The relative bonding ratio of other wells is given by the formula = (FP_sample - FP_free) / (FP_no_ The calculation was performed according to comp-FP_free. The relative binding rate and the concentration of the test polypeptide were calculated. The curve was plotted using this method, and the IC50 value was calculated by curve fitting.

[0055] The amino acid characteristics of SE-DR-bound polypeptides and unbound polypeptides are comprehensively considered. Then, P1, P4, P6, and P9 affect the affinity between polypeptides and SE-DR. It can be identified as an important amino acid. In particular, P1 is an amino acid with a hydrophobic side chain. It is an acid, and a rare or unnatural amino acid with similar properties; the P4 site is nonpolar, polar and amino acids that are uncharged, polar, and negatively charged, and rare or non-natural amino acids with similar properties. It is a natural amino acid; the P6 site is an amino acid with a short side chain, and is a rare amino acid with similar properties. It is a small or non-natural amino acid; the P9 site is an amino acid with a moderate to short side chain. These are rare or non-natural amino acids with similar properties.

[0056] [Table 1] (Note that P1-P9 in the table indicate the binding site of the polypeptide to HLA-DR. (Since cytoplasmic phosphate groups APL36 and APL37 are special, their binding sites are not shown.)

[0057] [Table 2] (Note that P1-P9 in the table indicate the binding sites of polypeptides to HLA-DR.)

[0058] Example 2: PBMC stimulation test Add 3 mL of Ficol to a 15 mL centrifuge tube, and then add 4 mL of blood sample to the Ficol. The solution was dropped onto the surface of the tube. The tube was centrifuged at 18-20°C with 400g for 30-40 minutes. Aspirate the upper layer of plasma with a Pasteur pipette, then aspirate the PBMC layer (approximately 2 mL), and at least The mixture was then added to a 14 mL centrifuge tube pre-filled with 3 volumes (6 mL) of PBS. The solution was gently resuspended. The tube was centrifuged at 18-20°C for 10 minutes at 60-100g. After cardiac arrest, remove the supernatant using a Pasteur pipette and suspend the resulting cell aggregate in 6-8 mL of PBS. Then, centrifuge at 18-20°C and 60-100°C for 10 minutes, and remove the supernatant. Add the cells to an appropriate amount. Resuspend in 640 complete medium, and after cell counting, 1 × 10⁶ 6 Diluted to cells / ml. The cell suspension was added to a 96-well plate. Allosteric peptide was added as a positive control. Tide was added to 100 μl of cell culture medium to achieve a final concentration of 10 μg / ml. The mixture was supplemented with 200 μl of 1640 complete medium. After 5 days of incubation, the supernatant was aspirated and 3 The mixture was centrifuged at 000 rpm for 5 minutes, and the IL-6 level in the supernatant was detected by ELISA. .

[0059] [Table 3]

[0060] Through Examples 1 and 2, we have examined several SE-DR affinity peptides at the animal level. Pharmacodynamic studies were conducted.

[0061] Part II: Pharmacodynamic Results of SE-DR Affinity Peptides 1. SE-DR affinity peptides significantly suppress disease progression in CIA rats. This promoted the differentiation of peripheral blood T cells in CIA rats into Th2 cells and Treg cells. Example 3) The collagen-induced arthritis (CIA) model is incomplete and involves type II bovine collagen (CII). Emulsions prepared using Freund's adjuvant (IFA) were used in female Lewis rats. The method was established by intradermal injection into the head and tail. After establishing the model, morphological changes in the limbs of rats were observed. We checked daily. When the inflammation score was 1, affected rats were used as a model group, and SE-DR affinity peptides were used. Groups (FNS007(APL2), APL7, APL9, APL20, APL21 respectively) (administered ALP37), autoantigen peptide group (i.e., type II collagen CII263- Participants were randomly divided into two groups: 272) and an unrelated peptide group. Polypeptide administration was performed on all participants except the model group. The dosage was set to 0.2 mmol in all cases. A control group was also established, and the drug was administered once every other day for 15 days. It was then injected into the tail vein. For efficacy analysis, the rats were administered once every other day starting from the day they were registered. The area under the curve (AUC) of the inflammation score was used until the administration was completed. Furthermore, after the completion of administration, 144 At a set time, blood is collected from the orbital angular vein, and helper T cells (Th1, Th2) are found in peripheral blood mononuclear cells. The proportion of Th17 cells and T regulatory cells (Treg) was measured by flow cytometry. .

[0062] As a result, SE-DR affinity peptides (FNS007(APL2), APL7, APL 9. Peripheral blood of rats that were repeatedly administered APL20, ALP21, and ALP37 for 144 hours. Table 4 shows the effect on the change in the proportion of T cell subtypes in the control group. In comparison, the proportion of Th1 and Th17 cells, and the proportion of Th1 / Th2 cells in the model group. The proportion of Th2 cells and Treg cells increased significantly (p<0.05, p<0.01), and the proportion of Th2 cells and Treg cells increased. The number of people with SE-DR affinity decreased significantly (p<0.05, p<0.01). Compared to the model group, SE-DR affinity was lower. In the group administered sex peptides, the proportion of Th1 and Th17 cells was significantly reduced (p<0.0). 5. The proportion of Th2 cells and Treg cells increased significantly (p<0.01). 5. p<0.01). The results are shown in Table 4. In conclusion, SE-DR affinity peptides are In CIA rats, inflammatory Th1 and Th2 cells in peripheral blood T cells were converted to anti-inflammatory Th This promoted differentiation into 2 and Treg cells.

[0063] [Table 4] Note: When compared to the control group. # P<0.05, ## P<0.01; compared with the model group. If * P<0.05, ** P<0.01.

[0064] 2. Test results of SE-DR affinity peptides in tuberculosis-positive RA patients 2.1: SE-DR affinity peptides do not inhibit normal immune function. SE-DR affinity peptides inhibit only the activation of T cells related to self-antigen recognition. Because it does not affect the normal immune system that protects the body from external infections, it reduces the risk of infections and tumors. It does not have [unclear] and can be used in tuberculosis-positive rheumatoid arthritis patients. Therefore, the present invention is applicable to tuberculosis-positive R A novel therapeutic agent can be provided for patient A, and anti-rheumatic biological agents of this type can be used. This solves the problem that it cannot be used in RA patients.

[0065] Studies have shown that SE-DR affinity peptides are effective in the immune responses of healthy humans, healthy monkeys, and healthy rats. It has been shown that it does not significantly alter the disease system, and that SE-DR affinity peptides are normal in the body. It does not interfere with immune function, does not reduce the body's defense capabilities, and does not pose a risk of infection or tumors. This suggests that... Please refer in particular to Examples 4 to 6 below.

[0066] Example 4 In SD rats, the following results were obtained after continuous administration of SE-DR affinity peptides for one month. The effects on immune indicators were investigated. SE-DR affinity peptide was administered to six groups (FNS00). Divided into 7(APL2), APL7, APL9, APL20, APL21, APL37), A control group was also established, with 10 males and 10 females in each group, who were administered the treatment once a day for 4 weeks. After the completion of administration... Anti-FNS007 antibodies in serum, various immunoglobulins in serum, complement in serum, peripheral blood T We analyzed lymphocytes and cytokines in serum.

[0067] As a result, after administration of SE-DR affinity peptide, all immunosuppressants were reduced compared to the control group. Since no significant changes were observed in the disease indicators, the SE-DR affinity peptide was found in healthy rats. It was suggested that it does not affect the immune system. Table 5 shows that SE-DR affinity peptides are effective in the immune system. The results of the effects on immune cells in rats are shown.

[0068] [Table 5]

[0069] Example 5 In a study using cynomolgus monkeys, the following results were obtained after continuous administration of SE-DR affinity peptides for one month. The effects on immune indicators in isal monkeys were investigated. The administration scheme for SE-DR affinity peptides was as follows: Six treatment groups (FNS007(APL2), APL7, APL9, APL20, ALP2) 1. Divide into groups ALP37), and set up a control group with 3 males and 3 females in each group, once a day. The drug was administered continuously for 4 weeks. After the completion of administration, the anti-FNS007 antibody in the serum and other serotype immunoglobulins were tested. Immune indicators such as globulin, serum complement, peripheral blood T lymphocytes, and serum cytokines are observed. I understand.

[0070] As a result, after administration, there were no significant changes in any immune indicator compared to the control group. No such effect was observed, indicating that SE-DR affinity peptides do not affect the immune system of healthy monkeys. Table 6 shows the effects of SE-DR affinity peptides on immune cells in healthy monkeys. vinegar.

[0071] [Table 6]

[0072] Example 6 The effects of a single dose of SE-DR affinity peptide on human immune indicators in healthy individuals. The study involved volunteers. The SE-DR affinity peptide used was FNS007. The administration method is divided into three dose groups (5, 10, and 20 mg / case), and placebo control A troll group was also established. After administration, different serotypes of immunoglobulins, peripheral blood T lymphocytes, and blood were tested. Cytokines and other factors in the patient were observed. As a result, after administration, the placebo group and the baseline group before administration were observed. Compared to the IN values, no significant changes were observed in any of the immune indicators, and SE-DR affinity peptide It was shown that the drug does not affect the immune system of healthy individuals. Table 7 shows the SE-DR affinity peptide. This shows the effect on immune cells in healthy individuals.

[0073] [Table 7]

[0074] 2.2: SE-DR affinity peptides do not pose a risk of inducing tuberculosis infection. Furthermore, we will compare the differences in infection induction between SE-DR affinity peptides and biological agents. Therefore, in this study, we used CIA model rats with latent tuberculosis infection and adalimumab and The incidence of tuberculosis during arthritis treatment with SE-DR affinity peptides was compared. As a result, Both dalimumab and SE-DR affinity peptides showed good therapeutic effects in CIA rats. However, adalimumab tends to induce tuberculosis recurrence during treatment, and the incidence of tuberculosis is high. The rate was high at 60%. On the other hand, in the model group and the SE-DR affinity peptide group, tuberculosis recurrence was It was not recognized. As described above, SE-DR affinity peptides are not effective against rheumatoid arthritis. It has good therapeutic effects, does not increase the tuberculosis infection rate, and is safe even for tuberculosis-positive RA patients. A certain outcome is expected. See Example 7 for details.

[0075] Example 7 Establishment of a latent infection model: 5 × 10⁶ female Lewis rats 3 E of CFU (Colony-forming Unit) The rats were infected intravenously with the rdman strain. The amount of Mycobacterium tuberculosis (MTB) in the lung and spleen tissue of the rats was: 1 x 10 times 4 weeks after infection 4 The above was reached. At this point, the anti-tuberculosis drug isoniazid (0 Rat lungs were subjected to the addition of 0.1g / L of rifampicin (15g / L) to drinking water for 4 weeks. Furthermore, tuberculosis bacteria cultures from the spleen became negative two weeks after discontinuation of the drug, confirming the latent tuberculosis infection model. This suggested that they had succeeded in establishing the position.

[0076] Using the female Lewis rats infected with latent tuberculosis described above, a latent tuberculosis infection CIA model was established. Method: Emulsion containing bovine CII and IFA (100 μg of collagen) was administered to rats infected with latent tuberculosis. The rats were immunized by intradermal injection of (containing g) into the head and tail. Seven days later, the rats were boosted once using the same method. Morphological changes in the limbs of rats were examined daily. Inflammation scores were set to 1 for the model group and SE-D. R affinity peptide group (FNS007(APL2), APL7, APL9, APL20, A Rats were subjected to a controlled environment in the LP21, ALP37, 0.2 mmol group and the adalimumab 1 mg / kg group. They were divided into groups of 10 rats each. The other 8 latent infected rats were not included in the model establishment. The control group was used. Both the model group and the control group were administered PBS. The appropriate medication is administered by tail vein injection at a volume of 5 ml / kg, once every other day for 21 consecutive days. The rats were administered. On the 22nd day after administration, the rats in each group showed signs of inflammation in their limbs. We evaluated A and detected MTB cultures from the lungs and spleen of rats at the time of sacrificing.

[0077] After the model was established, the inflammation score in the model rats was significantly increased compared to the control group. This reflects the influence of the drug on indicators throughout the entire pathological process and evaluates the overall effect of the drug. Therefore, the area under the curve (AUC) of the inflammation score was statistically analyzed. As a result, the model group In comparison, the SE-DR affinity peptide group and the adalimumab group showed a decrease in inflammation in affected rats. We were able to significantly reduce A (P<0.01). At the time of sacrifice, the control group, Cultures of Mycobacterium tuberculosis from the lungs and spleens of rats in the model group and the SE-DR affinity peptide group were all negative, but in the adalimumab group, cultures of Mycobacterium tuberculosis from the lungs and spleens were positive in 6 / 10 rats . Also, in the adalimumab group, obvious toxic symptoms such as piloerection and decreased activity were observed during administration, but the condition of rats after administration of SE-DR affinity peptide was good. From the above results, it was suggested that SE-DR affinity peptide did not increase the risk of tuberculosis recurrence in rats, and its safety was superior to that of adalimumab. The results are shown in Table 8. The results are shown in Table 8.

[0078]

Table 8

[0079] 2.3: SE-DR affinity peptide has no risk of inducing hepatitis B (HBV) infection Furthermore, to compare the differences in infection induction between SE-DR affinity peptide and biological agents, in this study, CIA mice, a model of chronic hepatitis B infection, were used, and the activation of hepatitis B in mice during arthritis treatment with abatacept and SE-DR affinity peptide was compared. The results showed that both abatacept and SE-DR affinity peptide showed good therapeutic effects on CIA mice, but abatacept tended to activate hepatitis B during the administration period,

[0080] and the ALT value increased to 100 IU / ml in 50% of the mice. On the other hand, SE-DR affinity peptide ​​No significant differences were observed in ALT levels among the peptide group, model group, and control group. This suggests that SE-DR affinity peptides reduce the activation rate of hepatitis B during treatment for rheumatoid arthritis. It is expected to be safe for use in RA patients infected with hepatitis B without increasing blood sugar levels. For details, please refer to Example 8.

[0081] Example 8 Establishment of a mouse model of chronic hepatitis B: Raav8-1.3H in DBA / 1 mice BV(1×10 12 A chronic hepatitis B (HBV) model was established by injecting (vg / mL) into the tail vein. Ta.

[0082] Using DBA / 1 mice infected with chronic hepatitis B, a chronic hepatitis B infection CIA model was developed. Established. Method: Raav8-1.3HBV (1×10⁻¹) was administered to mice. 12 (vg / mL) tail Two days after injection into the pulse, an emulsion of bovine CII and CFA (containing 100 μg of collagen) was administered. Mice were immunized by intradermal injection into the head and tail. After 21 days, the drug was prepared using bovine CII and IFA. The emulsion (containing 100 μg of collagen) was injected once into the peritoneal cavity of mice to boost their performance. Subsequently, morphological changes in the limbs of the mice were examined daily, and those with an inflammation score of 1 were randomly assigned to the treatment group. We identified the problem. We used diseased mice as the model group and SE-DR affinity peptide group (FNS007(APL 2) 50 μmol each of APL7, APL9, APL20, ALP21, and ALP37 (1 dose), then randomly divided into the abatacept group (100 μg), with 12 animals in each group. Ten mice with latent hepatitis type 1 infection were not used to establish the model and were instead used as a control group. The control group was administered PBS. The mice in each group received 10 ml / kJ of the drug corresponding to their size. The drug was administered by tail vein injection in a dose of g, once every other day for 27 consecutive days. On the 28th day after administration, Mau The sac was sacrificed. During administration, the inflammation score of the limbs of mice in each group was evaluated. Booster immunization On the day of (22 days after tail vein injection) and at the time of sacrifice, the virus injection of mice and liver function were examined. Serological indicators were detected.

[0083] After establishing the mouse model, the inflammation score of the model mice was significantly higher compared to the control group. It rose. Reflecting the impact of the drug on indicators throughout the entire course, the overall effect of the drug is evaluated. Therefore, the area under the curve (AUC) of the inflammation score was statistically analyzed. As a result, the model group and In comparison, the SE-DR affinity peptide group and the abatacept group showed improved heat irritation scores in affected mice. We were able to reduce it to a certain level (P<0.01).

[0084] In the evaluation of hepatitis B virus, during booster immunization, i.e., 22 days after tail vein injection: HBsAg, HBe antigen, and HBV DNA were detected in all mice, and ALT and AS were detected. The mean T values ​​were 45 and 65 U / L, respectively. This is for establishing a chronic infection model of hepatitis B. Success was suggested. At the point of sacrifice, the control group, model group and SE- Serological indicators of liver function in mice with DR affinity peptides were the same as those observed during booster immunization. However, serological indicators related to liver function in mice administered with abatacept were significantly higher. The levels rose, and in 50% of the mice, ALT levels rose to over 100 IU / ml, so This suggests a significant increase in the risk of hepatitis B activation in the Us. Furthermore, During batacept administration, obvious toxic symptoms such as decreased piloerection and activity were observed, but SE - The mice were in good condition after administration of the DR affinity peptide. These results are from SE-D R-affinity peptides did not increase the risk of hepatitis B activation in mice, and their safety was demonstrated by Avatar This suggests that it is superior to Sept. The results for each metric at the time of final sampling are shown in Table 9. Please refer to the following.

[0085] [Table 9] Note: When compared to the control group. # P<0.05, ## P<0.01; compared with the model group. If * P<0.05, ** P<0.01.

[0086] 3. Results of the study on combination therapy in RA patients The combination of antigen-specific and non-antigen-specific therapies represents the future direction of rheumatoid arthritis treatment. On the other hand, non-antigen-specific therapies facilitate antigen-specific therapies and allow for early control of clinical symptoms (such as inflammation). On the other hand, the immunomodulatory effect of antigen-specific drugs can reconstruct immune tolerance. The disease can be completely controlled.

[0087] To test this theory, this study uses the CIA model to investigate SE-DR affinity peptides. Tide (antigen-specific therapeutic agent) and adalimumab / abatacept / methotrexate (non-antigen-specific therapeutic agent). We are considering combination therapy with specific therapeutic agents, including SE-DR affinity peptide, adalimumab, and avatar. The differences were compared with Sept or methotrexate alone. As a result, SE-DR affinity peptide Combination therapy with tide and adalimumab, combination therapy with SE-DR affinity peptide and abatacept The combination therapy of the method and SE-DR affinity peptides and methotrexate is SE-DR affinity Compared to peptide monotherapy, it was shown to rapidly suppress the inflammatory response in rats.

[0088] Compared to adalimumab, abatacept, or methotrexate monotherapy, the combination therapy group showed more It was effective and safe. In the SE-DR affinity peptide group and the combination group, the condition persisted even after discontinuation of administration. The therapeutic effect was maintained, but in the adalimumab group, abatacept group, or methotrexate group, Inflammation scores increased after discontinuation of administration. This suggests that SE-DR affinity peptides and non-antigen-specific By using alternative therapies (adalimumab / abatacept / methotrexate, etc.) in combination Furthermore, it can significantly improve clinical remission rates and maintain remission even after discontinuation of the drug, thus enabling gradual tapering or intermediate use of the drug. This suggests that stopping the process can be achieved. See Examples 9, 10, and 11 for details.

[0089] Example 9 Female Lewis rats were used to establish a CIA model. Methods: Rats were given bovine CII and An emulsion prepared by IFA was administered intradermally to the head and tail to immunize the fish. Seven days later, the same method was used once more. The rats were boosted. Afterward, morphological changes in the rats' limbs were examined daily. (Model group, SE-D) R-affinity peptide group (FNS007, 0.2 mmol), adalimumab group (1 mg / kg ), and the combination therapy group (FNS0070.2 mmol + adalimumab 1 mg / kg) were among the three groups. They were divided into groups of 10 rats each. Another group of 8 female Lewis rats was excluded from the model establishment. The control group was used as the model group. Both the model group and the control group were administered PBS. The appropriate medication is administered via intravenous injection at a dose of 5 mL / kg, once every other day for 21 days. The drug was administered daily. During administration (once every other day), the inflammation score of the limbs of rats in each group was evaluated, and the drug was administered. The area under the curve (AUC) of inflammation scores during administration was calculated. The limbs of rats in each group were also examined during discontinuation of administration. The inflammation score was intermittently assessed (once every other day), and the rats were sacrificed 22 days after discontinuation of administration. The procedure was followed. HE staining of the ankle joint was performed to assess tissue damage. The specific scoring criteria are as follows: The scale is as follows: 0 = Normal synovial tissue; 1 = Synovial hyperplasia and inflammatory cell infiltration; 2 = Pannus formation and cartilage erosion; 3 = extensive destruction of cartilage accompanied by subchondral bone erosion; 4 = loss of joint integrity, a Kilosis.

[0090] As a result, in the CIA rat model, FNS007 performed better than the FNS007 monogroup. The combination of 7 and adalimumab was shown to rapidly suppress the inflammatory response in rats. This indicates that the inflammation score in the first week of administration was higher in the combination group compared to the FNS007 monotherapy group. This decline was demonstrated by a decrease in the AUC of the inflammation score, from 14.26 to 11.93. There was a significant difference between these two groups (P<0.05). The inflammation score AUC in the combination therapy group was the same for all administrations. During the period, there was a significant decrease, with the inflammation score decreasing from 46.17 to 35.88. These two groups There was a statistically significant difference between the two groups (P<0.05). Furthermore, compared to adalimumab, the combination therapy group The effectiveness was more pronounced (inflammation score decreased from 54.51 to 35.88, showing a statistically significant difference). (p<0.05), the incidence of infection in rats decreased, and symptoms of piloerection and epistaxis in rats were present. The condition improved significantly. At the same time, administration was discontinued on the 21st day, followed by a 21-day observation period. During discontinuation of administration, the FNS007 group and the combination therapy group were able to maintain their therapeutic effect, but adalimumab The group was unable to maintain the therapeutic effect, and this was due to a significant increase in inflammation scores compared to when the treatment was discontinued. This was also evident from the results (P<0.01). The results are shown in Table 10, Figures 1 and 2.

[0091] [Table 10] Note: When compared to the control group. # P<0.05, ## P<0.01; compared with the model group. If * P<0.05, ** P<0.01.

[0092] Example 10 Male DBA mice were used to establish the CIA model. Methods: Bovine CII and CFA (10 Mice were immunized by intradermal injection of an emulsion (containing 0 μg of collagen) into the head and tail. 21 days later, an emulsion of bovine CII and IFA (containing 100 μg of collagen) was injected intraperitoneally. The mice were injected and given one boost. Afterward, morphological changes in the mice's limbs were observed daily. At the point when the inflammation score reached 1, mice were randomly assigned to the treatment group. Affected mice were used as the model group, and SE-DR parents were used. Japanese peptide group (FNS007, 0.2 mmol), abatacept group (5 mg / kg), In the combination therapy group (FNS007, 0.2 mmol + abatacept, 5 mg / kg), there were no cases in the three groups. They were divided into groups of 12 mice each. Another group of 10 female DBA / 1 mice was used for model establishment. The control group was designated as the model group. Both the model group and the control group were administered PBS. The appropriate drug for the mice was administered via tail vein injection at a dose of 10 mL / kg, once every other day for 28 days. The drug was administered daily, followed by a 28-day recovery period. During administration (once every other day), the mice in each group... Inflammation scores in the limbs were evaluated, and the area under the curve (AUC) of the inflammation score during administration was calculated. During discontinuation of administration, the inflammation score of the limbs of rats in each group was continuously evaluated (once every other day), and administration was performed. Rats were sacrificed 28 days after discontinuation. HE staining of the ankle joint was performed to assess tissue damage. The specific scoring criteria are as follows: 0 = Normal synovial tissue; 1 = Synovial hyperplasia and inflammation. Symptomatic cell infiltration; 2 = Pannus formation and cartilage erosion; 3 = Major cartilage fracture accompanied by subchondral bone erosion. Destruction; 4 = Loss of joint integrity, ankylosis.

[0093] As a result, in the CIA mouse model, FNS00 was compared to the FNS007-only group. The combination therapy of 7 and abatacept has been shown to rapidly suppress the inflammatory response in mice. This indicates that, during the first 10 days of administration, compared to the FNS007 monotherapy group, Inflammation scores decreased in the combination therapy group, and the AUC of inflammation scores decreased from 25.64 to 18.11. This was demonstrated by the results. A significant difference was observed between the two groups (p<0.05). The inflammation score AUC decreased significantly from 90.23 to 75.87 throughout the entire administration period. A significant difference was observed between the groups (P<0.05). In addition, compared to the abatacept group, the combination therapy The effectiveness of the method group was more pronounced (inflammation score AUC decreased from 89.16 to 75.87). A statistically significant difference was observed (p<0.05), the incidence of infection in mice decreased, and symptoms of piloerection and epistaxis were reduced. The condition improved significantly. At the same time, administration was discontinued on day 28, followed by a 28-day observation period. During discontinuation of administration, the FNS007 group and the combination therapy group were able to maintain their therapeutic effect, but abatacept The treatment effect could not be maintained, and this was due to a significant increase in inflammation scores compared to when the treatment was discontinued. This was also evident from the results (P<0.01). The results are shown in Table 11.

[0094] [Table 11] Note: When compared to the control group. # P<0.05, ## P<0.01; compared with the model group. If * P<0.05, ** P<0.01.

[0095] Example 11 Female Lewis rats were used to establish the CIA model. Method: Emulsion prepared with bovine CII and IFA was injected intradermally into the tail base for immunization. Seven days later, the rats were boosted once by the same method. Thereafter, the morphological changes of the rats' limbs were observed daily, and rats with an inflammation score of 1 were randomly assigned to the treatment group. The diseased rats were randomly divided into three groups, namely the model group, the SE-DR affinity peptide group (APL20, 0.2 mmol), the methotrexate group (0.5 mg / kg), and the combination therapy group (APL20, 0.2 mmol + methotrexate , 0.5 mg / kg), with 10 rats in each group. Another 8 female Lewis rats were not used for model establishment and served as the control group. The model group and the control group were administered PBS. The corresponding drugs for the rats in each group were injected into the tail base at a dose of 5 mL / kg for administration. Methotrexate was administered twice a week, and the other drugs were administered once every other day for 21 consecutive days, and then the rats were allowed to recover for 21 days. The inflammation scores of the rats' limbs in each group were evaluated during administration (once every other day), and the area under the curve (AUC) of the inflammation scores during administration was calculated. During the period of suspension of administration, the inflammation scores of the rats' limbs in each group were continuously evaluated (once every other day). The rats were sacrificed on the 22nd day after the suspension of administration. For tissue injury evaluation, HE staining of the ankle joints was performed. The specific scoring criteria are as follows: 0 = normal synovial tissue; 1 = synovial hyperplasia and inflammatory cell infiltration; 2 = pannus formation and cartilage erosion; 3 = most destruction of cartilage with subchondral bone erosion; 4 = loss of joint integrity, ankylosis. As a result, in the CIA rat model, compared with the APL20 alone group, APL20 and

[0096] In the CIA rat model, compared with the APL20 alone group, APL20 and Combination therapy with methotrexate has been shown to rapidly suppress inflammatory responses in rats. This indicates that, in the first week of administration, the combination therapy was superior to that of the FNS007 monotherapy group. In the control group, inflammation scores decreased, and the inflammation score AUC decreased from 14.93 to 12.06. This was demonstrated. There was a significant difference between the two groups (P<0.05). Inflammation scores in the combination therapy group AUC decreased significantly from 48.69 to 37.59 throughout the entire administration period, and no significant difference was observed between the two groups. (P<0.05). Furthermore, the efficacy of the combination therapy group compared to methotrexate was This was more pronounced (the inflammation score AUC decreased from 67.69 to 37.59, a statistically significant difference). (p<0.05), symptoms of piloerection and epistaxis in rats were significantly improved, and animal mortality was reduced. Administration was discontinued on the 21st day, followed by a 21-day observation period. Among the groups, the APL20 group and the combination therapy group were able to maintain their treatment efficacy, but the methotrexate group was unable to treat the treatment. The effect could not be maintained. This was because the inflammation score was significantly higher compared to when the treatment was discontinued. This was also evident from the results (P<0.01). The results are shown in Table 12.

[0097] [Table 12] Note: When compared to the control group. # P<0.05, ## P<0.01; compared with the model group. If * P<0.05, ** P<0.01.

[0098] As described above, SE-DR affinity peptides and non-antigen-specific therapeutic agents (adalimumab / A) By combining it with batacept / methotrexate, etc., the clinical remission rate is significantly improved. This allows for the maintenance of remission even after discontinuation of the drug, and enables the gradual reduction or discontinuation of the drug. Therefore, the present invention can significantly improve clinical remission rates and reduce or moderate drug use. This allows us to provide a novel pharmaceutical composition that is expected to stop the disease.

[0099] Without departing from the spirit and scope of the present invention, various modifications and variations may be made in the present invention. It will be obvious to those skilled in the art that this is possible. Accordingly, the present invention is as described in the attached claims. Within the scope of the range and its equivalents, it is intended to include such modifications and alterations. Yes, they are.

Claims

1. The use of pharmaceutical compositions for the preparation of drugs for treating rheumatoid arthritis, The pharmaceutical composition comprises an SE-DR affinity peptide, a non-antigen-specific anti-rheumatic drug, and a pharmaceutically acceptable carrier. The aforementioned SE-DR affinity peptide is selected from FKGEQAGAGE, MGPKGRTVIIEQSWGSPKVTK, MGPKGRTVIIEQSLGSPKVTK, SIDLKDKKYKNIGAKLVQDVANNTNEEA, SIDLKDKKYKNIGAKLVQLVANNTNEEA, QYMCitADQAAGGLR, LTQCitGSVLR, WYNCitCHAAN, VETCitDGQVI, or VCitLCitSSVESTCitGRSCitPAPPPACitGLT. The aforementioned non-antigen-specific antirheumatic drug is adalimumab or abatacept. The aforementioned SE-DR affinity peptide competitively inhibits the binding of autoantigen peptides associated with rheumatic diseases to HLA-DR molecules, thereby regulating the disrupted immune microenvironment in the body and restoring immune balance. The pharmaceutical composition is characterized by its rapid inhibition of the progression of rheumatoid arthritis and its achievement of sustained remission even after discontinuation of the drug.

2. The use according to claim 1, characterized in that the pharmaceutically acceptable carrier comprises a binder, a surfactant, a solubilizer, a stabilizer, a lubricant, a wetting agent and / or a diluent.

3. The use according to claim 1, characterized in that the dosage form of the pharmaceutical composition is a capsule, tablet, pill, granule, elixir, ointment, patch, aerosol, injection, or suppository.

4. The use according to claim 1, characterized in that the dosage form of the pharmaceutical composition is a film-coated tablet, a sublingual preparation, a chewable preparation, a topical preparation, or an inhalation preparation.

5. The use according to claim 1, characterized in that the dosage form of the pharmaceutical composition is a transdermal formulation.

6. The use according to claim 1, wherein the pharmaceutical composition is suitable for various routes of administration, and the route of administration is selected from one or more of the following: oral administration, intravenous administration, subcutaneous administration, intramuscular administration, intracerebral administration, intranasal administration, intrapulmonary administration, intra-arterial administration, intra-articular administration, intradermal administration, intravitreous administration, intramedullary injection, intraperitoneal administration, intrathecal administration, or transdermal administration.

7. The use according to claim 1, characterized in that the patient with rheumatoid arthritis is tuberculosis-positive or tuberculosis-negative, and the patient with rheumatoid arthritis has or does not have hepatitis B.

Images