WO2023006125A1 - Se-dr亲和肽在制备治疗风湿疾病的药物中的用途 - Google Patents
Se-dr亲和肽在制备治疗风湿疾病的药物中的用途 Download PDFInfo
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- 229940126586 small molecule drug Drugs 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
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- 230000003637 steroidlike Effects 0.000 description 1
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- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 1
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/10—Peptides having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
Definitions
- the invention belongs to the field of biopharmaceuticals, and specifically relates to the use of SE-DR affinity peptides in the preparation of medicines for treating rheumatic diseases of tuberculosis-positive and/or rheumatic patients combined with hepatitis B.
- the present invention also relates to a pharmaceutical composition comprising SE-DR affinity peptide and non-antigen-specific anti-rheumatic disease medicine and its use in preparing medicine for treating various types of rheumatic diseases.
- Rheumatism is a group of extremely common clinical syndromes with arthralgia and fear of wind and cold as the main symptoms.
- Rheumatism is the abbreviation of rheumatic diseases, which generally refers to a large group of diseases affecting bones, joints, muscles and surrounding soft tissues, such as bursae, tendons, fascia, blood vessels, and nerves.
- Rheumatoid arthritis the most common rheumatic disease, is a chronic, systemic autoimmune disease characterized pathologically by erosive synovitis that can cause destruction of articular cartilage and bone, leading to joint deformities and eventually disabled.
- the three-year disability rate of patients without formal treatment is as high as 75%, which is one of the main reasons for the loss of young and middle-aged labor force.
- the average life expectancy of patients is shortened by 3-10 years compared with normal people, which is called "cancer of immortality".
- the disease has brought serious social and economic problems. 50% of patients who can still work at the early stage of the disease will lose their working ability after 10 years, and it will rise to 67% after 15 years.
- Rheumatoid arthritis is described abroad as "5D", that is, disease-causing, depression-causing, disability-causing, death-causing and bankruptcy-causing.
- the prevalence of rheumatoid arthritis in the world is 0.5%-1%, in my country it is about 0.3%, and the number of patients is nearly 5 million.
- the peak age of onset is 35-44 years old for female patients in China, and 55-64 years old for male patients, and the incidence rate increases with age. It is more common in women, and the incidence ratio of women to men is 2:1-3
- the pathogenesis of rheumatic diseases is very complex. Under the influence of environmental factors or genetic factors, the initial pathogenic antigen can activate the natural immune system in sensitive populations, and the antigen will activate T cells after being presented by antigen-presenting cells (APC).
- APC antigen-presenting cells
- the activation of T cells requires the co-stimulation of two independent signals.
- the first signal is the formation of a three-molecule complex of "human leukocyte antigen (HLA)-antigen peptide-T cell receptor (TCR)", in which HLA and antigen peptide first form two molecules complex, and then combined with TCR to form a trimolecular complex;
- the second signal is the combination of CD28 on the surface of T cells and CD80/86 on the surface of APC cells.
- Activated T cells can further activate B cells.
- T cells and B cells enter the blood circulation and reach the lesion sites containing self-antigens with similar structures to pathogenic antigens. Since self-antigens as pathogenic antigens can continuously activate T, B and other cells secrete a large amount of pro-inflammatory cytokines, chemokines and collagenases, which stimulate synovial hyperplasia and pannus formation, and continuous inflammation eventually leads to bone destruction.
- T lymphocytes play an important role in the initiation, inflammation persistence and relapse stages of rheumatoid arthritis, especially the differentiation of activated T cells into helper T cells. increasingly prominent. The change of Th17 cell and Treg cell level has become a research hotspot in this field.
- drugs for the treatment of rheumatic diseases can be divided into four generations based on the time and principle of development: the first generation is non-steroidal anti-inflammatory drugs (NSAIDs); the second generation is glucocorticoids (GC) ; The third generation is a disease-modifying drug (slow-acting antirheumatic drug) (DMARD); the fourth generation is a biological agent represented by a TNF- ⁇ inhibitor.
- the fourth-generation drugs for the treatment of rheumatic diseases include TNF- ⁇ inhibitors, IL-1 antagonists, IL-6 antagonists, JAK3 inhibitors, T cell inhibitors and B cell inhibitors, etc., currently in the US, Japan and Europe markets The proportion of consumption is as high as 97%.
- the currently commonly used therapeutic drugs for rheumatic diseases are not enough in terms of safety and effectiveness.
- Methotrexate which is recommended as a first-line clinical drug, has a median lethal dose of 43 mg/kg in rats, and its main toxicity is bone marrow suppression and gastrointestinal toxicity.
- the preclinical toxicity of etanercept in clinical second-line biological agents is mainly reflected in latent tuberculosis infection, and its clinical toxicity is mainly the induction of immune diseases, severe infections and increased cancer incidence; the preclinical safety carcinogenicity experiment of abatacept Among them, about 50% of the mice in the administration group died from lymphoma, and the incidence of breast cancer also showed an increase.
- tuberculosis For patients with a positive tuberculin test, treatment for tuberculosis is required before the use of biological agents, and even if the tuberculin test is negative, active tuberculosis needs to be monitored during the application of biological agents; for patients with hepatitis B
- prophylactic antiviral therapy should be given
- HBcAb-positive/HBsAg-negative patients start non-rituximab
- HBcAb-positive/HBsAg-negative patients start non-rituximab
- RA rheumatic diseases
- NSAIDs, GCs and DMARDs have extensive immunosuppressive effects and cannot specifically inhibit lymphocytes related to the pathogenesis; biological agents and new small molecule drugs are targeted drugs for RA, but most of them are aimed at the middle and lower reaches of the pathogenesis of RA Cytokines and kinases, while inhibiting the above factors, have brought side effects, such as the inhibition of TNF- ⁇ , which can lead to an increase in the incidence of tuberculosis infection and tumor;
- TNF- ⁇ inhibitors and DMARDs its target is still the activation of a wide range of T cells, rather than disease-specific T cells, which can cause immune deficiency while treating diseases, leading to severe infections , increasing the incidence of cancer. Therefore, the development of RA therapeutic drugs targeting disease-specific targets is the main way to solve the safety problems
- SE-DR affinity peptide (for example, FNS007) is an innovative drug independently developed by my country that directly targets T lymphocytes. It can be said to be the fifth-generation drug for the treatment of rheumatic diseases (especially RA).
- SE-DR affinity peptide plays a role by competitively inhibiting the combination of rheumatic disease-associated antigen peptides and major histocompatibility complex molecules, and can interfere with the first signal of rheumatic disease-associated antigen-mediated specific T cell activation "HLA -The formation of antigenic peptide-T lymphocyte receptor (TCR)"trimolecular complex can inhibit the activation and proliferation of disease-specific autoreactive T cells, change the differentiation of T cells and the secretion of downstream cytokines, and achieve the treatment of rheumatic diseases (especially is the purpose of RA).
- SE-DR affinity peptides are allosteric peptides of the main autoantigen epitopes of RA, which act on the specific links of the occurrence and development of RA, and can Inhibit the combination of RA-related various antigenic peptides and HLA molecules, and regulate the disordered immune microenvironment in patients, reduce the number of inflammatory cells, increase the number of cells that inhibit inflammation, and through long-term correction, it is expected to rebuild immune tolerance , completely prevent the progression of the disease. It can be said that the mechanism of action of SE-DR affinity peptides (for example, FNS007) in the treatment of rheumatic diseases represents the development direction of anti-rheumatic drugs in the future.
- FNS007 is a Class 1.1 new drug developed by Hebei Phoenix Biotechnology Co., Ltd. It has entered the clinical research stage and is conducting phase I clinical research.
- the present invention is an in-depth study on the new indications of SE-DR affinity peptides (especially, FNS007) aimed at the defects in the safety and effectiveness of the existing drugs for treating rheumatic diseases. Therefore, an object of the present invention is to provide the use of SE-DR affinity peptide in the preparation of medicines for treating rheumatic diseases in tuberculosis-positive and/or rheumatic patients combined with hepatitis B.
- Another object of the present invention is to provide a pharmaceutical composition comprising SE-DR affinity peptide and a non-antigen-specific anti-rheumatic disease drug and its use in the preparation of drugs for treating various types of rheumatic diseases.
- Antigen-specific therapy is a brand-new idea to solve the clinical shortage of drugs for the treatment of rheumatic diseases. Although some non-antigen-specific therapeutic drugs (such as TNF- ⁇ inhibitors) have achieved some effects, how to induce the body's tolerance to self-antigens is the key to the treatment of autoimmune diseases, and more specific The target also makes it possible to reduce drug toxicity such as infection. Therefore, antigen-specific therapy, or its combined therapy with non-antigen specificity has become a new trend in the treatment of rheumatic diseases.
- the present invention provides the use of SE-DR affinity peptide in the preparation of medicines for treating rheumatic diseases in tuberculosis positive and/or patients with rheumatic diseases combined with hepatitis B, said SE-DR affinity peptide Refers to peptides that bind to shared epitopes of HLA DR molecules.
- the shared epitope is the 70-74th amino acid of the HLA-DR molecule, both of which express the QK/RRAA sequence, and the SE-DR affinity peptide competitively inhibits the antigenic peptide associated with rheumatic diseases and the HLA DR molecule
- the combination of these drugs plays a role, but it does not have a wide range of immunosuppressive effects, and there is no risk of inducing infection or tumor.
- the core sequence of the SE-DR affinity peptide combined with SE-DR includes core amino acids corresponding to P1 to P9, wherein the P1 site is an amino acid containing a hydrophobic side chain and has similar properties
- the rare or unnatural amino acids, the P4 position is non-polar, polar uncharged, polar negatively charged amino acids and rare or unnatural amino acids with similar properties.
- the corresponding P1 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from Tyr (Y), Phe (F), Trp (W), Leu (L), Ile (I ), Met (M), Val (V) or Ala (A), the P4 site is selected from Met (M), Ala (A), Val (V), Ile (I), Leu ( One of L), Asp(D), Glu(E), Gln(Q), Ser(S) or Cit.
- the corresponding P1 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from one of Tyr (Y), Phe (F) or Trp (W), and the P4 The site is selected from one of Met (M), Leu (L), Asp (D), Glu (E) or Cit.
- the corresponding P6 site in the core sequence of the SE-DR affinity peptide and SE-DR is an amino acid containing a short side chain and a rare or unnatural amino acid with similar properties
- the P9 site is for amino acids with medium and short side chains and rare or unnatural amino acids with similar properties.
- the corresponding P6 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from Ala (A), Gly (G), Ser (S), Thr (T) or Asn (N ), the P9 site is selected from one of Ala (A), Gly (G), Leu (L) or Met (M).
- the SE-DR affinity peptide comprises the following amino acid sequence: FXGEQGXXGE, or FXGEXAXXGE, wherein: X is selected from P, K, Q, A or G.
- X is selected from A or G. More preferably, the SE-DR affinity peptide is FNS007 (FKGEQAGAGE).
- the SE-DR affinity peptide comprises the following amino acid sequence: YXKQXTXXLA, wherein: X is selected from V, A, G, N, L or K.
- X is selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide is selected from PKYVKQNTLKLAT, PGYVKQGTLGLAT, YVKQNTLKLA, YVAQNTLKLA, YAKQATLKLA or YAKQATLALA.
- the SE-DR affinity peptide comprises the following amino acid sequence: IWYIXCFXCEXHXXL, wherein: X is selected from A, G, M, T, V, N, Q or S.
- X is selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide is selected from IWYINCFGCETHAML, IWYIQCFGCETHAML, IWYISCFGCETHAML, IWYITCFGCETHAML, IWYINCFACETHAML or IWYINCFVCETHAML.
- the SE-DR affinity peptide comprises the following amino acid sequence: RSFXLAXSXXGVG, wherein: X is selected from A, G, T, S or E.
- X is selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide is selected from RSFTLASSETGVG, RSFALASSETGVG, RSFTAASSETGVG, RSFTLDSSETGVG, RSFTLAASETGVG, RSFTLASSATGVG, RSFTLASSEAGVG or RSFTLDSSETGVG.
- the SE-DR affinity peptide comprises the following amino acid sequence: SAVXLCitXSXXGVR, wherein: X is selected from A, G, R, S, V or P.
- the amino acid sequence of the SE-DR affinity peptide is SAVRLCitSSVPGVR, SAVELCitSSVPGVR, SAVDLCitSSVPGVR, SAVGLCitSSVPGVR, SAVRLCitFSVPGVR, SAVRLCitSSVEGVR, SAVRLCitSSVKGVR, SAVRLCitSSVWGVR, SAVRLCitWSVPGVR, SAVRLCitSSVRGVR, SAVRLCitKSVPGVR, SAVALCitSSVPGVR.
- the SE-DR affinity peptide comprises the following amino acid sequence: GVYXTCitXSXXCitLCit, wherein: 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.
- the SE-DR affinity peptide comprises the following amino acid sequence: QDXNCitXNXXKNS, wherein: 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 is QDFTNCitANKLKNS.
- the SE-DR affinity peptide comprises the following amino acid sequence: VVLLVATXGCitXRXXSAYQDK, wherein: X is selected from A, G, E, V or N. Preferably, X is selected from A or G. More preferably, the amino acid sequence of the SE-DR affinity peptide is VVLLVATEGCitVRVNSAYQDK.
- the rheumatic disease is selected from rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, undifferentiated spondyloarthropathy, systemic lupus erythematosus, systemic sclerosis or one or more of collagen diseases.
- the present invention provides a pharmaceutical composition for treating rheumatic diseases, the pharmaceutical composition comprising SE-DR affinity peptide and non-antigen-specific anti-rheumatic disease drugs, and a pharmaceutically acceptable carrier , preferably, the weight ratio of the SE-DR affinity peptide to the non-antigen-specific anti-rheumatic disease drug is 50:1-1:50, more preferably, the weight ratio can be selected from 20:1-1: 20, 15:1-1:15, 10:1-1:10, 5:1-1:5, 4:1-1:4, 3:1-1:3 or 2:1-1:2,
- the specific dosage ratio is related to the type of non-antigen-specific anti-rheumatic disease drugs selected, and can be determined by clinicians.
- the SE-DR affinity peptide refers to the peptide segment that binds to the shared epitope of the HLA DR molecule.
- the shared epitope is the 70-74th amino acid of the HLA-DR molecule, both of which express the QK/RRAA sequence, and the SE-DR affinity peptide competitively inhibits the antigenic peptide associated with rheumatic diseases and the HLA DR molecule.
- the combination of the drugs can improve the disordered immune microenvironment in the patient's body and restore the immune balance.
- the pharmaceutical composition quickly inhibits the progress of rheumatic diseases and achieves sustained relief after the drug is reduced or stopped.
- the core sequence of the SE-DR affinity peptide combined with SE-DR includes core amino acids corresponding to P1 to P9, wherein the P1 site is an amino acid containing a hydrophobic side chain and has Rare or unnatural amino acids of similar properties.
- the corresponding P1 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from Tyr (Y), Phe (F), Trp (W), Leu (L), Ile (I ), Met (M), Val (V) or Ala (A); the P4 site is non-polar, polar uncharged, polar negatively charged amino acids and rare amino acids with similar properties or unnatural amino acids.
- the corresponding P4 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from Met (M), Ala (A), Val (V), Ile (I), Leu (L ), Asp(D), Glu(E), Gln(Q) or Ser(S).
- the corresponding P6 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is an amino acid containing a short side chain and a rare or unnatural amino acid with similar properties
- the P9 site is a medium- and short-chain-containing amino acid.
- Amino acids with side chains and rare or unnatural amino acids with similar properties Preferably, the corresponding P6 site in the core sequence of the SE-DR affinity peptide combined with SE-DR is selected from Ala (A), Gly (G), Ser (S), Thr (T) or Asn (N ), the P9 site is selected from one of Ala (A), Gly (G), Leu (L) or Met (M).
- the non-antigen-specific anti-rheumatic disease drugs are selected from drugs targeting cytokines, B-cell targets, T-cell targets, kinase targets, traditional disease-modifying anti-rheumatic drugs (DMARDs), non-steroidal One or more of anti-inflammatory drugs (NSAIDs) or hormonal drugs.
- the non-antigen-specific anti-rheumatic disease drugs are selected from tumor necrosis factor (TNF) inhibitors, IL-6 inhibitors, IL-1 inhibitors, IL-17 inhibitors, RANKL inhibitors, T cell co-stimulators
- TNF tumor necrosis factor
- IL-6 inhibitors IL-6 inhibitors
- IL-1 inhibitors IL-17 inhibitors
- RANKL inhibitors T cell co-stimulators
- signaling inhibitors CD20 inhibitors, CD22 inhibitors, BLyS and APRIL inhibitors, BAFF inhibitors, JAK inhibitors, BTK inhibitors, Syk inhibitors, IRAK4 inhibitors, p38 inhibitors or small molecule immunosuppressants or more.
- the non-antigen-specific anti-rheumatic disease drug is selected from adalimumab or its biosimilars, tocilizumab or its biosimilars, anakinra or its biosimilars, tofacitinib One or more of nicillin or its biosimilars, abatacept or its biosimilars, rituximab or its biosimilars, methotrexate or leflunomide.
- the SE-DR affinity peptide comprises the following amino acid sequence: FXGEQGXXGE, or FXGEXAXXGE, wherein: X is selected from K, Q, A or G. Preferably, X is selected from A or G. More preferably, the SE-DR affinity peptide is FNS007 (FKGEQAGAGE).
- the amino acid sequence of the SE-DR affinity peptide is selected from the group consisting of: MGPKGRTVIIEQSWGSPKVTK, MGPKGRTVIIEQSLGSPKVTK, SIDLKDKKYKNIGAKLVQDVANNTNEEA, SIDLKDKKYKNIGAKLVQLVANNTNEEA, QYMCitADQAAGGLR, LTQCitGSVLR, WYNCitAPPCHAAN, VEPPitDGQVISTPAPCitLCGitRSS.
- the pharmaceutically acceptable carrier includes a binder, a surfactant, a solubilizer, a stabilizer, a lubricant, a wetting agent and/or a diluent, and the pharmaceutical composition
- the dosage forms of the drug are capsules, tablets, pills, liquids, powders, granules, fine granules, film-coated agents, precipitants, troches, sublingual agents, chewing agents, buccal agents, pastes, syrups , suspensions, elixirs, emulsions, coatings, ointments, plasters, dressings, percutaneous preparations, lotions, inhalants, aerosols, injections or suppositories.
- the pharmaceutical composition is suitable for administration by various routes, including oral, intravenous, subcutaneous, intramuscular, intracerebral, intranasal, pulmonary, intraarterial, intraarticular , intradermal, intravitreal, intraosseous infusion, intraperitoneal, intrathecal or transdermal administration.
- the rheumatic disease is selected from rheumatoid arthritis, juvenile idiopathic arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, undifferentiated spondyloarthropathies , one or more of systemic lupus erythematosus, systemic sclerosis, collagen disease, Crohn's disease, and ulcerative colitis.
- the patient with rheumatic disease is tuberculosis positive or negative; the patient with rheumatic disease is combined with hepatitis B or not combined with hepatitis B.
- the present invention has the following beneficial effects:
- the antigen-specific therapeutic drug SE-DR affinity peptide (especially, FNS007) had no significant effect on the immune system of healthy people, healthy cynomolgus monkeys and healthy rats.
- SE-DR affinity peptide especially, FNS007
- CIA type II collagen-induced arthritis model
- SE-DR affinity peptides especially, FNS007
- CIA type II collagen-induced arthritis
- antigen-specific therapeutic drugs especially, FNS007, APL20
- non-antigen-specific therapeutic drugs such as adalimumab, abatacept, or methotrexate
- Antigen-specific therapeutic drugs can be used in combination with other non-antigen-specific therapeutic drugs to achieve the goal of completely controlling rheumatoid arthritis.
- Figure 1 In the experiment shown in Example 9, the inflammation scores at various time points during administration of the CIA rat model.
- Figure 2 In the experiment shown in Example 9, the inflammation scores at various time points during drug withdrawal in the CIA rat model.
- test materials used in the following examples are commercially available products unless otherwise specified.
- the polypeptide used was synthesized by solid phase synthesis and purified by high performance liquid chromatography by Hebei Phoenix Biotechnology Co., Ltd.
- the result of mass spectrometry analysis of the polypeptide shows that the sequence of the polypeptide is correct and the purity is above 95%.
- SE-DR molecules are directly related to RA pathogenesis. It can present RA-related autoantigens in vivo, such as type II collagen (CII), heat shock protein 60 (Hsp60), cartilage glycoprotein (HCgp-39), etc., and then activate autoreactive T cells, leading to rheumatoid arthritis happened.
- An important strategy for the treatment of RA is to use competitive inhibition to block the activation of self-reactive T cells by self-antigen peptides.
- the current main idea is to modify the prototype peptide so that it can inhibit the binding of self-antigens and not activate self-reactive T cells. This requires allosteric peptide molecules to have a certain affinity for SE-DR and show Decreased activation of T cells. Based on the above theoretical basis, we designed and synthesized a series of allosteric peptides, and screened their affinity with SE-DR and the activation ability of peripheral blood T cells of RA patients. See Examples 1 and 2 for specific steps and results.
- reaction system 96-well plate, 200 ⁇ L of reaction system per well, containing 500 nM HLA-DR1, 25 nM fluorescently labeled peptides and serially diluted peptides to be tested, starting from 200 ⁇ M concentration, 5-fold dilution 5 gradients, the last well does not contain the peptide to be tested Peptides were tested as quality control wells. Quality control wells use fluorescently labeled peptides without MHCII molecules to measure free peptide signals.
- the pH of the reaction system was 7.4, and protease inhibitor cocktail was added before use, and incubated at 37°C for 30min.
- P1, P4, P6 and P9 are the key amino acids that affect the affinity of the polypeptide and SE-DR.
- the properties of P1 are amino acids with hydrophobic side chains and rare or unnatural amino acids with similar properties;
- P4 sites are non-polar, polar uncharged, polar negatively charged amino acids and rare or unnatural amino acids with similar properties.
- P6 sites are amino acids with short side chains and rare or unnatural amino acids with similar properties;
- P9 sites are amino acids with medium and short side chains and rare or unnatural amino acids with similar properties.
- P1-P9 in the table indicates the binding site of the polypeptide and HLA-DR, due to the particularity of the APL36 and APL37 polypeptides, the binding position is not indicated
- Embodiment 2 PBMC stimulation test
- Example 1 we conducted animal-level pharmacodynamic studies on some of the SE-DR affinity peptides.
- SE-DR affinity peptide can significantly inhibit the disease progression of CIA rats, and promote the differentiation of peripheral blood T cells of CIA rats to Th2 and Treg cells (Example 3)
- the emulsion prepared with bovine type II collagen (CII) and incomplete Freund's adjuvant (IFA) was injected intradermally at the root of the tail of female Lewis rats, and boosted once 7 days later in the same way to establish a collagen-induced arthritis model (Collagen- induced arthritis, CIA).
- CII bovine type II collagen
- IFA incomplete Freund's adjuvant
- a blank control group was also set up, and the drug was injected into the tail vein once every other day for 15 consecutive days.
- the inflammation score was started on the day when the rats were enrolled in the group, once every other day, until the end of the administration, and the curative effect was analyzed by using the area under the curve (AUC) of the inflammation score.
- AUC area under the curve
- blood was collected from the orbital inner canthus vein 144 hours after the end of the administration, and then flow cytometry was used to measure the proportion of helper T cells (Th1, Th2, Th17) and regulatory T cells (Treg) in peripheral blood mononuclear cells .
- the SE-DR affinity peptide only inhibits the activation of T cells related to self-antigen recognition, and has no effect on the body's normal immune system against foreign infections, it does not have the risk of infection and tumors, and can be used for tuberculosis-positive patients. Rheumatoid patients, so the present invention can provide a brand-new therapeutic drug that can be used for tuberculosis-positive RA patients, and solves the problem that anti-rheumatic biological products cannot be used in this type of RA patients.
- SE-DR affinity peptides have no significant changes in the immune systems of healthy people, healthy monkeys and healthy rats, suggesting that SE-DR affinity peptides do not interfere with the body's normal immune function and will not reduce the body's defense capabilities. No risk of infection and tumors. For details, see the following Examples 4-Example 6.
- SE-DR affinity peptides were divided into 6 dose groups (FNS007(APL2), APL7, APL9, APL20, ALP21, ALP37), and a blank control group was set up, with 10 female and 10 male animals in each group, administered daily 1 time, continuous administration for 4 weeks.
- FNS007(APL2), APL7, APL9, APL20, ALP21, ALP37 6 dose groups
- a blank control group was set up, with 10 female and 10 male animals in each group, administered daily 1 time, continuous administration for 4 weeks.
- serum binding antibodies, different types of serum immunoglobulins, serum complement, analysis of peripheral blood T lymphocytes and serum cytokines were observed.
- Cynomolgus monkeys were used to investigate the effect of continuous administration of SE-DR affinity peptide for one month on the immune indexes of cynomolgus monkeys.
- SE-DR affinity peptides were divided into 6 dose groups (FNS007(APL2), APL7, APL9, APL20, ALP21, ALP37), and a blank control group was set up, with 3 female and 3 male animals in each group, administered daily 1 time, continuous administration for 4 weeks.
- serum binding antibodies, different types of serum immunoglobulins, serum complement, analysis of peripheral blood T lymphocytes and serum cytokines were observed.
- SE-DR affinity peptide was FNS007, which was divided into 3 dose groups (5, 10 and 20 mg/case), and a placebo control group was set up. After administration, different types of serum immunoglobulins, peripheral blood T lymphocytes were analyzed and analyzed. Serum cytokines and other indicators were observed.
- this study used a CIA rat model with tuberculosis latent infection to compare the effects of adalimumab and SE-DR affinity peptides on the treatment of arthritis in rats. In, the incidence of tuberculosis in rats.
- CFU Mycobacterium tuberculosis
- the above-mentioned female Lewis rats latently infected with tuberculosis were used to establish a CIA model of latent tuberculosis infection.
- Rats with disease were randomly divided into model group, SE-DR affinity peptide group (respectively given FNS007(APL2), APL7, APL9, APL20, ALP21, ALP37, dose 0.2mmol) and adalimumab 1mg/kg group, each group 10 only.
- 8 latently infected rats without modeling were set as the blank group.
- the model group and the blank group were given PBS, and the rats in each group were injected with the corresponding drugs according to the tail vein of the volume of 5ml/kg, once every other day. After 21 days of administration, the rats were sacrificed on the 22nd day after administration. During the administration period, the inflammation scores of the four paws of the rats in each group were evaluated, and the MTB culture of the rats' lungs and spleens were detected when they were sacrificed.
- the inflammation score of the modeled rats was significantly increased.
- Area (AUC) was used for statistical analysis. The results showed that, compared with the model group, both the SE-DR affinity peptide group and the adalimumab administration group could significantly reduce the inflammation score of the diseased rats (P ⁇ 0.01).
- Table 8 The therapeutic effect of SE-DR affinity peptide on CIA rats latently infected with tuberculosis and the observation of tuberculosis recurrence
- this study used the CIA mouse model of chronic hepatitis B infection to compare the effects of abatacept and SE-DR affinity peptides in the treatment of arthritis in mice , HBV activation in mice.
- mice DBA/1 mice were used to inject Raav8-1.3HBV (1 ⁇ 10 12 vg/mL) virus into mice through tail vein to establish HBV chronic hepatitis B model.
- mice with chronic hepatitis B infection were used to establish a CIA model of chronic hepatitis B infection.
- mice were randomly divided into model group, SE-DR affinity peptide group (administered FNS007 (APL2), APL7, APL9, APL20, ALP21, ALP37 respectively, dose 50 ⁇ mol) and abatacept group (dose 100 ⁇ g), each group 12 Only.
- FNS007 APL2
- APL9 APL7
- ALP21 ALP37
- abatacept group dose 100 ⁇ g
- 10 latent hepatitis B-infected mice without modeling were set as the blank group.
- the model group and the blank group were given PBS, and the mice in each group were given the corresponding drug by tail vein injection at a volume of 10ml/kg, once every other day.
- mice After administration for 27 days, the mice were sacrificed on the 28th day after administration. During the administration period, the inflammation scores of the four paws of the mice in each group were evaluated; on the day of booster immunization (22 days after the tail vein injection of the virus) and when the samples were finally sacrificed, the serological indexes of the mice were detected for virus serology and liver function.
- the inflammation score of the modeled mice was significantly increased.
- Area (AUC) was used for statistical analysis. The results showed that compared with the model group, both the SE-DR affinity peptide group and the abatacept group could significantly reduce the inflammation score of the diseased mice (P ⁇ 0.01).
- mice were boosted immunized that is, 22 days after the tail vein injection of the virus, all mice could be detected positive for HBsAg, HBeAg, and HBV DNA, and the average levels of ALT and AST were 45 and 65 U/L, respectively. It is suggested that the hepatitis B chronic infection model has been prepared successfully.
- the serum indexes of liver function of the mice in the blank group, the model group and the SE-DR affinity peptide group were at the same level as those in the booster immunization, but the serum indexes of liver function in the abatacept group were significantly increased, and 50% of the mice The ALT level in the mice increased to >100IU/ml, suggesting that the risk of hepatitis B activation in the mice was greatly increased.
- the animals had obvious toxic symptoms such as piloerection and decreased activity, while the mice were in good condition after the administration of the SE-DR affinity peptide.
- SE-DR affinity peptide does not increase the risk of hepatitis B activation in mice, and its safety is better than that of abatacept. See Table 9 for the results of each index when the final materials were collected.
- non-antigen-specific therapy can assist antigen-specific therapy to control clinical symptoms (such as inflammation) as soon as possible, and on the other hand, through the immunomodulatory effect of antigen-specific drugs, immune tolerance can be rebuilt and the disease can be completely controlled.
- this study adopts the CIA model to investigate the combined application of SE-DR affinity peptide (antigen-specific therapeutic drug) and adalimumab/abatacept/methotrexate (non-antigen-specific therapeutic drug).
- SE-DR affinity peptide antigen-specific therapeutic drug
- adalimumab/abatacept/methotrexate non-antigen-specific therapeutic drug
- SE-DR affinity peptides and non-antigen-specific therapeutic drugs can greatly improve the clinical remission rate, and the remission can be sustained after drug withdrawal, achieving Reduce the drug withdrawal. See embodiment 9, embodiment 10 and embodiment 11 for details.
- the CIA model was established by using female Lewis rats.
- Model group SE-DR affinity peptide group (using FNS007, 0.2mmol), adalimumab group (1mg/kg), and combined drug group (FNS007 0.2mmol+adalimumab Monoclonal antibody 1mg/kg), 10 rats in each group.
- 8 female Lewis rats without modeling were set as the blank group, the model group and the blank group were given PBS, and the rats in each group were injected with the corresponding drugs according to the volume of 5mL/kg tail vein, once every other day. Medicine for 21 days, recovery for 21 days after drug withdrawal.
- the inflammation scores of the four paws of the rats in each group were evaluated (once every other day), and the area under the curve (AUC) of the inflammation scores during the administration period was calculated.
- AUC area under the curve
- the inflammation scores of the four paws of the rats in each group once every other day
- the rats on the 22nd day after drug withdrawal take the ankle joints for HE staining, and evaluate the tissue damage.
- mice Male DBA/1 mice were used to establish the CIA model.
- METHODS Intradermal injection of the emulsion (containing 100 ⁇ g collagen) prepared from bovine CII and CFA was used to immunize the base of the tail of the mice, and a booster immunization was given by intraperitoneal injection of the emulsion prepared from bovine CII and IFA (containing 100 ⁇ g collagen) 21 days later. After that, the morphological changes of the four paws of the mice were checked daily, and when the inflammation score was 1, they were randomly enrolled into the group and given treatment.
- mice with the disease were randomly divided into three groups, namely: model group, SE-DR affinity peptide group (using FNS007, 0.2mmol), abatacept group (5mg/kg), and combined drug group (FNS007 0.2mmol+abatacept 5mg/kg), 12 rats in each group.
- model group SE-DR affinity peptide group (using FNS007, 0.2mmol), abatacept group (5mg/kg), and combined drug group (FNS007 0.2mmol+abatacept 5mg/kg), 12 rats in each group.
- Another 10 unmodeled female DBA/1 mice were set as the blank group, the model group and the blank group were given PBS, and the mice in each group were given the corresponding drugs by tail vein injection at a volume of 10 mL/kg, once every other day. Continuous administration for 28 days, recovery for 28 days after drug withdrawal.
- the inflammation scores of the four paws of the mice in each group were evaluated (once every other day), and the area under the curve (AUC) of the inflammation scores during the administration period was calculated.
- AUC area under the curve
- the drug withdrawal observation period continue to evaluate the inflammation scores of the four paws of the mice in each group (once every other day), and execute the mice on the 28th day after the drug withdrawal, and take the ankle joints for HE staining to evaluate the tissue damage.
- the CIA model was established by using female Lewis rats.
- Diseased rats were randomly divided into three groups, namely: model group, SE-DR affinity peptide group (using APL20, 0.2mmol), methotrexate group (0.5mg/kg), and combined drug group (APL20 0.2mmol+ formazan aminopterin 0.5mg/kg), 10 rats in each group.
- mice 8 female Lewis rats without modeling were set as the blank group.
- the model group and the blank group were given PBS.
- the rats in each group were given the corresponding drugs by tail vein injection at a volume of 5 mL/kg, and methotrexate was administered weekly. 2 times, other medicines were administered once every other day, for 21 consecutive days, and recovered for 21 days after drug withdrawal.
- the inflammation scores of the four paws of the rats in each group were evaluated (once every other day), and the area under the curve of the inflammation scores during the administration period was calculated.
- 0 points normal synovial tissue
- 1 point synovial hyperplasia, inflammatory cell infiltration
- 2 points pannus formation, cartilage erosion
- 3 points most cartilage destruction with subchondral bone erosion
- 4 points joint integrity Loss, joint stiffness.
- the combination of SE-DR affinity peptide group and non-antigen-specific therapeutic drugs can greatly improve the clinical remission rate, and can Sustained remission, to achieve drug withdrawal. Therefore, the present invention can provide a brand-new pharmaceutical composition that can significantly improve the clinical remission rate and is expected to reduce drug withdrawal.
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Abstract
提供了SE-DR(含有共享表位的HLA-DR分子)亲和肽在制备治疗结核阳性和/或合并乙型肝炎的风湿疾病患者的风湿疾病的药物中的用途,解决了这种类型的风湿疾病患者药品选择受限的难题。还提供了一种包含SE-DR亲和肽和非抗原特异性抗风湿疾病药物的药物组合物及其在制备治疗各种类型的风湿疾病的药物中的用途。所述药物组合物能够快速抑制风湿疾病的进程,并且能够在减停药之后实现持续缓解,达到了彻底控制风湿疾病的目的。
Description
相关申请的交叉引用
本申请要求2021年07月30日提交的中国申请号202110873116.6的权益。所述申请号202110873116.6据此全文以引用方式并入本文。
本发明属于生物制药领域,具体涉及SE-DR亲和肽在制备治疗结核阳性和/或合并乙型肝炎的风湿疾病患者的风湿疾病的药物中的用途。另外,本发明还涉及一种包含SE-DR亲和肽和非抗原特异性抗风湿疾病药物的药物组合物及其在制备治疗各种类型的风湿疾病的药物中的用途。
风湿疾病是以关节痛、畏风寒为主症的一组极其常见的临床症候群。风湿病是风湿疾病的简称,泛指影响骨、关节、肌肉及其周围软组织,如滑囊、肌腱、筋膜、血管、神经等一大组疾病。类风湿关节炎(RA)是一种最常见的风湿疾病,这是一种慢性、全身性自身免疫疾病,病理特征为侵蚀性滑膜炎,可造成关节软骨和骨质破坏,引起关节畸形最终致残。未经正规治疗的患者3年致残率高达75%,是造成中青年劳动力丧失的主要原因之一。患者的平均寿命较正常人缩短3-10年,被称为“不死的癌症”。该疾病带来了严重的社会及经济问题,发病初期仍可工作的患者,10年后有50%丧失工作能力,15年后将升至67%。国外将类风湿关节炎描述为“5D”,即致病、致抑郁、致残、致死和致破产。世界范围内类风湿关节炎的患病率为0.5%-1%,我国约为0.3%,患者人数近500万。国内女性患者发病高峰年龄在35-44岁,男性患者为55-64岁,随年龄增长发病率增加。女性多见,女性与男性发病率比例为2:1-3:1。
风湿疾病的发病机制非常复杂,敏感人群在环境因素或遗传因素的影响下,初始致病抗原可激活天然免疫系统,抗原经抗原递呈细胞(APC)递呈后激活T细胞。T细胞的激活需要两个独立信号协同刺激,第一信号为形成“人白细胞抗原(HLA)-抗原肽-T细胞受体(TCR)”三分子复合物,其中HLA与抗原肽先形成二分子复合物,再与TCR结合形成三分子复合物;第二个信号为T细胞表面的CD28与和APC细胞表面的CD80/86结合。被激活的T细胞可进一步激活B细胞,这些激活的T细胞、B细胞进入血液循环,到达含有与致病抗原类似结构的自身抗原的病灶部位,由于自身抗原作为致病抗原可不断激活T、B等细胞,分泌大量的致炎性细胞因子、趋化因子及胶原酶类,刺激滑膜增生、血管翳形成,炎症持续发生最终导致骨破坏。T淋巴细胞在类风湿关节炎的启动、炎症持续和复发阶段均起重要作用,特别是激活后的T细胞向辅助性T细胞分化,在类风湿关节炎的发病机制及治疗药物的研究领域地位日益凸显。Th17细胞及Treg细胞水平的变化已成为该领域的研究热点。
目前,治疗风湿疾病(特别是RA)的药物依据发展的时间及原理,主要可以分为四代:第一代是非甾体类抗炎药物(NSAID);第二代是糖皮质激素(GC);第三代是改善病情药(慢作用抗风湿药)(DMARD);第四代是以TNF-α抑制剂为代表的生物制剂。第四代治疗风湿疾病的药物包括TNF-α抑制剂、IL-1拮抗剂、IL-6拮抗剂、JAK3抑制剂、T细胞抑制剂和B细胞抑制剂等等,目前在美日欧市场的用量占比高达97%。但是,目前临床上常用的风湿疾病(特 别是RA)的治疗药物在安全性及有效性方面都存在不足。
目前常用的NSAID、GC、DMARD及生物制剂类药物均存在某些安全性问题,给临床应用带来一定的局限性。作为临床一线推荐用药的甲氨蝶呤,其在大鼠中的半数致死剂量为43mg/kg,主要的毒性为骨髓抑制和消化道毒性。临床二线生物制剂中的依那西普临床前的毒性主要体现为潜伏性结核菌感染,其临床毒性主要为诱发免疫疾病、严重感染和增加癌症发生率;阿巴西普的临床前安全性致癌实验中,给药组约有50%小鼠的死亡原因是淋巴癌,乳腺癌的发生率也显示提高。临床试验表明阿巴西普同样能够加重患者感染及提高罹患癌症的几率。多数生物制剂(如TNF抑制剂)FDA都给予了黑框警告,提示肿瘤和感染风险,患者在用药前需要进行结核菌素试验和乙型肝炎相关检查。对于结核菌素试验阳性的患者,需要先针对结核进行治疗后才能应用生物制剂,并且即便是结核菌素试验阴性,也需要在应用生物制剂的过程中监测活动性结核病;对于合并乙型肝炎的患者,HBcAb阳性患者开始利妥昔单抗治疗,或HBsAg阳性患者开始任何生物或靶向合成DMARD治疗时,需给予预防性抗病毒治疗;HBcAb阳性/HBsAg阴性患者开始非利妥昔单抗的生物或靶向合成DMARD治疗时,密切监测乙肝情况。因此,对于结核阳性或者合并乙型肝炎的患者,存在药物选择受限以及用药风险增加的问题。
此外,目前临床常用药物以不同的治疗方案治疗及不同的临床标准在治疗两年后随访,全球范围内的缓解率仅能达到40%左右,仍有近一半的患者无法得到有效的治疗。在我国,现有药物治疗后以不同的临床标准随访,RA患者的临床缓解率仅为8.6%-25.2%。有相当数量的患者无法通过目前的药物治疗获得临床缓解和病情控制。
现有临床上能够缓解风湿疾病(特别是RA)的治疗药物存在安全性问题的共同原因可以归结为“作用靶点特异性不足”。NSAID、GC及DMARD,具有广泛的免疫抑制作用,不能特异性抑制与发病相关的淋巴细胞;生物制剂及新型小分子药物虽然为RA的靶向药物,但大部分是针对RA发病机制中下游的细胞因子及激酶,在抑制上述因子的同时带来了副作用,如抑制TNF-α后可导致结核感染及肿瘤发生率的增加;阿巴西普是针对RA发病最上游的T细胞激活发挥作用的,虽然其安全性优于TNF-α抑制剂及DMARD,但其作用靶点仍是针对广泛的T细胞激活,而非疾病特异性T细胞,在治疗疾病的同时可引起免疫缺陷,进而导致严重感染,增加癌症的发生率。因此,开发针对疾病特异性靶点的RA治疗药物,是解决现有药物安全性问题的主要途径。
SE-DR亲和肽(例如,FNS007)是直接针对T淋巴细胞发挥作用的我国自主研发的创新药物,可以说是治疗风湿疾病(特别是RA)的第五代药物。SE-DR亲和肽通过竞争性抑制风湿疾病相关的抗原肽与主要组织相容性复合体分子的结合发挥作用,能够干扰风湿疾病相关抗原介导的特异性T细胞激活的第一信号“HLA-抗原肽-T淋巴细胞受体(TCR)”三分子复合体的形成,抑制疾病特异性自身反应性T细胞的活化和增殖,改变T细胞分化及下游细胞因子分泌,达到治疗风湿疾病(特别是RA)的目的。“抗原特异性治疗”是SE-DR亲和肽作用机制的一大特点,SE-DR亲和肽是RA主要自身抗原表位的变构肽,作用于RA发生和发展的特异性环节,能够抑制RA相关的多种抗原肽与HLA分子的结合,并且调节患者体内紊乱的免疫微环境,降低致炎性的细胞数量,升高抑制炎症的细胞数量,通过长期的纠正,有望重建免疫耐受,彻底阻止疾病的进展。可以说,SE-DR亲和肽(例如,FNS007)治疗风湿疾病的作用机制代表了未来抗风湿药物的发展方向。
FNS007是河北菲尼斯生物技术有限公司主导开发的一个1.1类新药,目前已进入临床研究阶段,正在开展I期临床研究。本发明是该公司针对现有的治疗风湿疾病的药物在安全性 和有效性方面存在的缺陷对SE-DR亲和肽(特别是,FNS007)新适应证的深入研究。因此,本发明的一个目的是提供SE-DR亲和肽在制备治疗结核阳性和/或合并乙型肝炎的风湿疾病患者的风湿疾病的药物中的用途。本发明的另一个目的是提供一种包含SE-DR亲和肽和非抗原特异性抗风湿疾病药物的药物组合物及其在制备治疗各种类型的风湿疾病的药物中的用途。
发明内容
对于现有风湿疾病的治疗药物来说,如何在临床安全性与有效性间找到平衡是关键。在一定范围内增加剂量可以使药物疗效增加、提高缓解率,但同时使药物的不良反应发生率增高。寻找更具疾病特异性的风湿疾病治疗药物及使用药物过程中的相关风险,是解决现有风湿疾病药物缓解率不足、寻找适应我国风湿疾病患者治疗需求的主要途径和全新尝试。
抗原特异性治疗是解决上述风湿疾病治疗药物临床不足的全新思路。尽管目前已有的一些非抗原特异性治疗药物(如TNF-α抑制剂)已经取得了一些效果,但如何诱导机体对自身抗原产生耐受,才是自身免疫性疾病治疗的关键,而更特异的靶点也让降低感染等药物毒性反应成为可能。因此,抗原特异性治疗,或其与非抗原特异性的联合治疗已成为风湿疾病治疗的新趋势。
为了实现本发明的目的,提供了以下技术方案:
在第一个方面中,本发明提供了SE-DR亲和肽在制备治疗结核阳性和/或合并乙型肝炎的风湿疾病患者的风湿疾病的药物中的用途,所述SE-DR亲和肽是指与HLA DR分子的共享表位结合的肽段。优选地,所述共享表位为HLA-DR分子的第70-74位氨基酸,均表达QK/RRAA序列,所述SE-DR亲和肽通过竞争性抑制风湿疾病相关的抗原肽与HLA DR分子的结合发挥作用,但不具备广泛的免疫抑制作用,无诱发感染、肿瘤的风险。
作为可选方式,在上述用途中,所述SE-DR亲和肽与SE-DR结合的核心序列包含P1至P9对应的核心氨基酸,其中P1位点为含疏水侧链的氨基酸及有类似性质的稀有或非天然氨基酸,P4位点为非极性、极性不带电荷、极性带负电荷的氨基酸及有类似性质的稀有或非天然氨基酸。
优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)、Trp(W)、Leu(L)、Ile(I)、Met(M)、Val(V)或Ala(A)中的一种,所述P4位点选自Met(M)、Ala(A)、Val(V)、Ile(I)、Leu(L)、Asp(D)、Glu(E)、Gln(Q)、Ser(S)或Cit中的一种。
更优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)或Trp(W)中的一种,所述P4位点选自Met(M)、Leu(L)、Asp(D)、Glu(E)或Cit中的一种。
作为可选方式,在上述用途中,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点为含短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,P9位点为含中、短侧链的氨基酸及有类似性质的稀有或非天然氨基酸。
优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点选自Ala(A)、Gly(G)、Ser(S)、Thr(T)或Asn(N)中的一种,所述P9位点选自Ala(A)、Gly(G)、Leu(L)或Met(M)中的一种。
作为可选方式,在上述用途中,所述SE-DR亲和肽包含下述氨基酸序列:FXGEQGXXGE,或FXGEXAXXGE,其中:X选自P、K、Q、A或G。
优选地,X选自A或G。更优选地,所述SE-DR亲和肽是FNS007(FKGEQAGAGE)。
作为可选方式,在上述用途中,所述SE-DR亲和肽包含下述氨基酸序列:YXKQXTXXLA,其中:X选自V、A、G、N、L或K。
优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列选自PKYVKQNTLKLAT、PGYVKQGTLGLAT、YVKQNTLKLA、YVAQNTLKLA、YAKQATLKLA或YAKQATLALA。
作为可选方式,在上述用途中,所述SE-DR亲和肽包含下述氨基酸序列:IWYIXCFXCEXHXXL,其中:X选自A、G、M、T、V、N、Q或S。
优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列选自IWYINCFGCETHAML、IWYIQCFGCETHAML、IWYISCFGCETHAML、IWYITCFGCETHAML、IWYINCFACETHAML或IWYINCFVCETHAML。
作为可选方式,在上述用途中,所述SE-DR亲和肽包含下述氨基酸序列:RSFXLAXSXXGVG,其中:X选自A、G、T、S或E。
优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列选自RSFTLASSETGVG、RSFALASSETGVG、RSFTAASSETGVG、RSFTLDSSETGVG、RSFTLAASETGVG、RSFTLASSATGVG、RSFTLASSEAGVG或RSFTLDSSETGVG。
作为可选方式,在上述用途中,所述SE-DR亲和肽包含下述氨基酸序列:SAVXLCitXSXXGVR,其中:X选自A、G、R、S、V或P。
优选地,所述SE-DR亲和肽的氨基酸序列为SAVRLCitSSVPGVR、SAVELCitSSVPGVR、SAVDLCitSSVPGVR、SAVGLCitSSVPGVR、SAVRLCitFSVPGVR、SAVRLCitSSVEGVR、SAVRLCitSSVKGVR、SAVRLCitSSVWGVR、SAVRLCitWSVPGVR、SAVRLCitSSVRGVR、SAVRLCitKSVPGVR、SAVALCitSSVPGVR或SAVRLCitRSVPGVR。
或者,所述SE-DR亲和肽包含下述氨基酸序列:GVYXTCitXSXXCitLCit,其中:X选自A、G或V。优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列为GVYATCitSSAVCitLCit。
或者,所述SE-DR亲和肽包含下述氨基酸序列:QDXNCitXNXXKNS,其中:X选自A、G、F、I、K或L。优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列为QDFTNCitANKLKNS。
或者,所述SE-DR亲和肽包含下述氨基酸序列:VVLLVATXGCitXRXXSAYQDK,其中:X选自A、G、E、V或N。优选地,X选自A或G。更优选地,所述SE-DR亲和肽的氨基酸序列为VVLLVATEGCitVRVNSAYQDK。
作为可选方式,在上述用途中,所述SE-DR亲和肽的氨基酸序列选自:MGPKGRTVIIEQSWGSPKVTK、MGPKGRTVIIEQSLGSPKVTK、SIDLKDKKYKNIGAKLVQDVANNTNEEA、SIDLKDKKYKNIGAKLVQLVANNTNEEA、QYMCitADQAAGGLR、LTQCitGSVLR、WYNCitCHAAN、VETCitDGQVI或VCitLCitSSVESTCitGRSCitPAPPPACitGLT。
作为可选方式,在上述用途中,所述风湿疾病选自类风湿关节炎、银屑病、银屑病关节炎、强直性脊柱炎、未分化脊柱关节病、系统性红斑狼疮、系统性硬化或胶原病中的一种或多种。
在第二个方面中,本发明提供了一种治疗风湿疾病的药物组合物,所述药物组合物包含SE-DR亲和肽和非抗原特异性抗风湿疾病药物,以及药学上可接受的载体,优选地,所述SE-DR亲和肽与非抗原特异性抗风湿疾病药物的重量比为50:1-1:50,更优选地,所述重量比可以选自20:1-1:20、15:1-1:15、10:1-1:10、5:1-1:5、4:1-1:4、3:1-1:3或2:1-1:2,其具体的用量比例与所选择的非抗原特异性抗风湿疾病药物的种类相关,可以由临 床医师决定,所述SE-DR亲和肽是指与HLA DR分子的共享表位结合的肽段,优选地,所述共享表位为HLA-DR分子的第70-74位氨基酸,均表达QK/RRAA序列,所述SE-DR亲和肽通过竞争性抑制风湿疾病相关的抗原肽与HLA DR分子的结合发挥作用,改善患者体内紊乱的免疫微环境,重建免疫平衡,所述药物组合物快速抑制风湿疾病的进程,并且在减停药之后实现持续缓解。
作为可选方式,在上述药物组合物中,所述SE-DR亲和肽与SE-DR结合的核心序列包含P1至P9对应的核心氨基酸,其中P1位点为含疏水侧链的氨基酸及有类似性质的稀有或非天然氨基酸。
优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)、Trp(W)、Leu(L)、Ile(I)、Met(M)、Val(V)或Ala(A)中的一种;所述P4位点为非极性、极性不带电荷、极性带负电荷的氨基酸及有类似性质的稀有或非天然氨基酸。优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P4位点选自Met(M)、Ala(A)、Val(V)、Ile(I)、Leu(L)、Asp(D)、Glu(E)、Gln(Q)或Ser(S)中的一种。所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点为含短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,所述P9位点为含中、短侧链的氨基酸及有类似性质的稀有或非天然氨基酸。优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点选自Ala(A)、Gly(G)、Ser(S)、Thr(T)或Asn(N)中的一种,所述P9位点选自Ala(A)、Gly(G)、Leu(L)或Met(M)中的一种。
所述非抗原特异性抗风湿疾病药物选自针对细胞因子靶点、针对B细胞靶点、针对T细胞靶点、针对激酶靶点的药物、传统改善病情的抗风湿药(DMARD)、非甾体抗炎药(NSAID)或激素类药物的一种或多种。
优选地,所述非抗原特异性抗风湿疾病药物选自肿瘤坏死因子(TNF)抑制剂、IL-6抑制剂、IL-1抑制剂、IL-17抑制剂、RANKL抑制剂、T细胞共刺激信号抑制剂、CD20抑制剂、CD22抑制剂、BLyS及APRIL抑制剂、BAFF抑制剂、JAK抑制剂、BTK抑制剂、Syk抑制剂、IRAK4抑制剂、p38抑制剂或小分子免疫抑制剂的一种或多种。
更优选地,所述非抗原特异性抗风湿疾病药物选自阿达木单抗或其生物类似物、托珠单抗或其生物类似物、阿那白滞素或其生物类似物、托法替尼或其生物类似物、阿巴西普或其生物类似物、利妥昔单抗或其生物类似物、甲氨蝶呤或来氟米特的一种或多种。
作为可选方式,在上述药物组合物中,所述SE-DR亲和肽包含下述氨基酸序列:FXGEQGXXGE,或FXGEXAXXGE,其中:X选自K、Q、A或G。优选地,X选自A或G。更优选地,所述SE-DR亲和肽是FNS007(FKGEQAGAGE)。
或者,所述SE-DR亲和肽的氨基酸序列选自:MGPKGRTVIIEQSWGSPKVTK、MGPKGRTVIIEQSLGSPKVTK、SIDLKDKKYKNIGAKLVQDVANNTNEEA、SIDLKDKKYKNIGAKLVQLVANNTNEEA、QYMCitADQAAGGLR、LTQCitGSVLR、WYNCitCHAAN、VETCitDGQVI或VCitLCitSSVESTCitGRSCitPAPPPACitGLT。
作为可选方式,在上述药物组合物中,所述药学上可接受的载体包括粘合剂、表面活性剂、增溶剂、稳定剂、润滑剂、湿润剂和/或稀释剂,所述药物组合物的剂型为胶囊剂、片剂、丸剂、液剂、散剂、颗粒剂、细粒剂、膜衣剂、沉淀剂、含片剂、舌下剂、咀嚼剂、颊剂、糊剂、糖浆剂、悬浮剂、酏剂、乳剂、涂布剂、软膏剂、硬膏剂、敷剂、经皮吸收型制剂、洗剂、吸入剂、气雾剂、注射剂或栓剂。
作为可选方式,在上述药物组合物中,所述药物组合物适于通过多种途径施用,包括口服、静脉内、皮下、肌内、脑内、鼻内、肺部、动脉内、关节内、皮内、玻璃体内、骨内输注、腹膜内、鞘内或透皮施用。
作为可选方式,在上述药物组合物中,所述风湿疾病选自类风湿关节炎、青少年特发性关节炎、银屑病、银屑病关节炎、强直性脊柱炎、未分化脊柱关节病、系统性红斑狼疮、系统性硬化、胶原病、克罗恩病、溃疡性结肠炎的一种或多种。
优选地,所述风湿疾病的患者是结核阳性的或结核阴性的;所述风湿疾病的患者是合并乙型肝炎的或者不合并乙型肝炎的。
本发明相对于现有技术,具有以下有益效果:
(1)首次发现抗原特异性治疗药物SE-DR亲和肽(特别是,FNS007)对健康人、健康食蟹猴和健康大鼠的免疫系统无明显影响。
(2)首次发现SE-DR亲和肽(特别是,FNS007)在潜伏感染结核杆菌的Ⅱ型胶原蛋白诱导性关节炎模型(CIA)大鼠中应用,与阿达木单抗相比,显著降低了结核发生率。
(3)首次发现SE-DR亲和肽(特别是,FNS007)在感染慢性乙肝的Ⅱ型胶原蛋白诱导性关节炎模型(CIA)小鼠中应用,与阿巴西普相比,显著降低了乙肝激活率。
(4)首次发现抗原特异性治疗药物(特别是,FNS007、APL20)与非抗原特异性治疗药物(如阿达木单抗、阿巴西普、或甲氨蝶呤)联合应用,可以快速抑制CIA大鼠疾病进展,且较单独使用阿达木单抗、阿巴西普或甲氨蝶呤安全性好,尤其是减停药之后可以持续缓解。
(5)抗原特异性治疗药物(如FNS007、APL20)可以与其他非抗原特异性治疗药物联合应用,达到彻底控制类风湿的目标。
图1:在实施例9所示实验中,CIA大鼠模型给药期间各时间点的炎症评分。
图2:在实施例9所示实验中,CIA大鼠模型停药期间各时间点的炎症评分。
下面参照具体的实施例对本发明做进一步说明。应当理解,此处所描述的具体实施例仅用于解释本发明,并不用于限定本发明的范围。
实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可通过正规渠道购买得到的常规产品。
下面实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为市售产品。
在本发明中,使用的多肽由河北菲尼斯生物技术有限公司以固相合成法合成、高效液相层析法纯化得到。所述多肽质谱分析结果显示多肽的序列正确,纯度在95%以上。
第一部分:SE-DR亲和肽的体外实验结果
SE-DR分子与RA发病直接相关。其在体内可递呈RA相关自身抗原,如Ⅱ型胶原(CII)、热休克蛋白60(Hsp60)、软骨糖蛋白(HCgp-39)等,进而激活自身反应性T细胞,导致类风湿关节炎的发生。治疗RA的一个重要策略就是利用竞争性抑制作用阻断自身抗原肽对自身反应性T细胞的活化。目前的主要思路是对原型肽进行改造,使其既能抑制自身抗原的结合, 又不会活化自身反应性T细胞,这就要求变构肽分子对SE-DR具有一定的亲和力,又表现出T细胞活化能力的降低。基于以上理论基础,我们设计合成理论一系列变构肽,对其与SE-DR的亲和力和RA病人外周血T细胞的活化能力进行了筛选,具体步骤和结果见实施例1和实施例2。
实施例1:多肽亲和力测定
配置反应体系:96孔板,每孔反应体系200μL,包含500nM HLA-DR1,25nM荧光标记多肽和梯度稀释的待测多肽,从200μM浓度开始,5倍稀释5个梯度,最后一孔不含有待测多肽,作为质控孔。质控孔采用不含MHCII分子的荧光标记多肽,用于测量游离多肽信号。反应体系pH7.4,用前加入蛋白酶抑制剂cocktail,37℃孵育30min。读取荧光偏振数值,将游离标记多肽读数平均值设定为FP_free,不加竞争肽的读数为FP_no_comp。计算其它孔的相对结合率数值,计算公式=(FP_sample-FP_free)/(FP_no_comp–FP_free)。利用相对结合率和待测多肽浓度制作曲线,通过拟合曲线计算IC
50。
综合SE-DR结合和非结合多肽的氨基酸性质,可以确定P1、P4、P6和P9是影响多肽和SE-DR的亲和的关键氨基酸。其中P1的性质为含疏水侧链的氨基酸及有类似性质的稀有或非天然氨基酸;P4位点为非极性、极性不带电荷、极性带负电荷的氨基酸及有类似性质的稀有或非天然氨基酸;P6位点为含短侧链的氨基酸及有类似性质的稀有或非天然氨基酸;P9位点为含中、短侧链的氨基酸及有类似性质的稀有或非天然氨基酸。
表1:通过拟合曲线计算的SE-DR结合多肽的IC
50
(注:表中P1-P9标明多肽和HLA-DR的结合位点,由于APL36和APL37多肽的特殊性,未对其结合位置进行标明)
表2:通过拟合曲线计算的非SE-DR结合多肽的IC
50
(注:表中P1-P9标明多肽和HLA-DR的结合位点)
实施例2:PBMC刺激试验
在15mL离心管中加入3mL ficoll,将4mL血液样品上样在ficoll表面。18-20℃400 g离心30-40min。将上层血浆用巴氏管吸出,将PBMC所在层(大致2mL)吸出,加到提前加入至少3倍体积(6mL)PBS的14mL离心管中,轻柔重悬。18-20℃下60-100g离心10min。离心后用巴氏管去除上清,所得细胞团用6-8mL PBS重悬,18-20℃60-100g离心10min,去上清。用适量的1640完全培养基重悬,细胞计数后稀释至1×10
6个/ml。将稀释过的细胞悬液加入96孔板。将阳性对照、变构肽加入100μl细胞培养液中,使终浓度为10μg/ml。用1640完全培养基补足至200μL。培养5天后,吸出上清,3000转/分钟,离心5分钟后ELISA检测上清IL-6的水平。
表3:ELISA检测上清IL-6的水平
通过实施例1和实施例2,我们对其中部分SE-DR亲和肽进行了动物水平药效研究。
第二部分:SE-DR亲和肽的药效学实验结果
1.SE-DR亲和肽能够显著抑制CIA大鼠疾病进展,并促进CIA大鼠外周血T细胞向Th2和Treg细胞分化(实施例3)
使用牛II型胶原(CII)同弗氏不完全佐剂(IFA)配制的乳剂,于雌性Lewis大鼠尾根皮内注射,7天后同法加强免疫1次,建立胶原诱导关节炎模型(Collagen-induced arthritis,CIA)。造模后每日检查大鼠四爪形态变化,炎症评分为1分时随机入组给予治疗,发病大鼠随机分为模型组、SE-DR亲和肽组(分别给予FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37)、原型肽(即,II型胶原CII263-272)组、无关肽组,除模型组外,多肽药物的给药剂量均为0.2mmoL。另设空白对照组,尾静脉注射给药,隔天一次,连续15天。从大鼠入组当天开始炎症评分,隔天一次,直至给药结束,采用炎症评分曲线下面积(AUC)进行疗效分析。另外,于给药结束后144h眼眶内眦静脉取血,其后采用流式细胞仪测定外周血单个核细胞中辅助性T细胞(Th1、Th2、Th17)及调节性T细胞(Treg)的比例。
结果显示,SE-DR亲和肽(FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37)多次 给药144h后对大鼠外周血T细胞亚型细胞比例变化的影响如表4所示:与空白组比较,模型组Th1、Th17细胞比例及Th1/Th2细胞比值明显升高(p<0.05,p<0.01),Th2、Treg细胞比例明显降低(p<0.05,p<0.01);与模型组比较,SE-DR亲和肽组Th1、Th17细胞比例明显降低(p<0.05,p<0.01),Th2、Treg细胞比例明显升高(p<0.05,p<0.01)。结果见表4。由此可见,SE-DR亲和肽促进CIA大鼠外周血T细胞由致炎性Th1和Th2向抑炎性的Th2和Treg细胞分化。
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
2.SE-DR亲和肽用于结核阳性RA对象的实验结果
2.1 SE-DR亲和肽不干扰机体正常的免疫功能
由于SE-DR亲和肽仅抑制与自身抗原识别相关的T细胞激活,而对机体的对抗外来感染的正常免疫系统无任何影响,因而其不存在感染和肿瘤的风险,可以用于结核阳性的类风湿患者,因此本发明能够提供一种可以用于结核阳性RA患者的全新治疗药物,解决了无法在这种类型的RA患者中使用抗风湿性生物制品的难题。
研究表明,SE-DR亲和肽对健康人、健康猴和健康大鼠的免疫系统均无明显改变,提示SE-DR亲和肽不干扰机体正常的免疫功能,不会降低机体的防御能力,无感染和肿瘤的风险。具体见以下实施例4-实施例6。
实施例4
采用SD大鼠,考察SE-DR亲和肽连续给药一个月对大鼠免疫指标的影响。SE-DR亲和肽分为6个剂量组(FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37),另设1个空白对照组,每组雌、雄各10只动物,每天给药1次,连续给药4周。给药结束进行血清结合抗体、血清不同类型免疫球蛋白、血清补体、外周血T淋巴细胞分析及血清细胞因子等指标观察。
结果表明,SE-DR亲和肽给药后,与空白组相比,各项免疫指标均无明显改变,提示SE-DR亲和肽对健康大鼠的免疫系统无影响。表5为SE-DR亲和肽对健康大鼠免疫细胞的影响结果。
表5:SE-DR亲和肽对大鼠免疫细胞的影响
实施例5
采用食蟹猴,考察SE-DR亲和肽连续给药一个月对食蟹猴免疫指标的影响。SE-DR亲和肽分为6个剂量组(FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37),另设1个空白对照组,每组雌、雄各3只动物,每天给药1次,连续给药4周。给药结束进行血清结合抗体、血清不同类型免疫球蛋白、血清补体、外周血T淋巴细胞分析及血清细胞因子等指标观察。
结果表明,SE-DR亲和肽给药后,与空白组相比,各项免疫指标均无明显改变,提示SE-DR亲和肽对健康猴的免疫系统无影响。表6为SE-DR亲和肽对健康猴免疫细胞的影响结果。
表6:SE-DR亲和肽对猴免疫细胞的影响
实施例6
采用健康受试者,考察SE-DR亲和肽单次给药对人体免疫指标的影响。SE-DR亲和肽采用FNS007,分为3个剂量组(5、10和20mg/例),另设安慰剂对照组,给药后进行血清不同类型免疫球蛋白、外周血T淋巴细胞分析及血清细胞因子等指标观察。
结果表明,SE-DR亲和肽给药后,与安慰剂组及给药前基线相比,各项免疫指标均无明显改变,提示SE-DR亲和肽对健康人体的免疫系统无影响。表7为SE-DR亲和肽对健康人免疫细胞的影响结果。
表7:SE-DR亲和肽对健康人免疫细胞的影响
2.2 SE-DR亲和肽无诱发结核感染的风险
为了进一步对比SE-DR亲和肽与生物制剂在诱发感染方面的差异,本研究采用结核潜伏感染的CIA大鼠模型,对比阿达木单抗和SE-DR亲和肽在治疗大鼠关节炎过程中,大鼠的结核发生率。
结果表明,阿达木单抗和SE-DR亲和肽均对CIA大鼠具有良好的治疗作用,但是阿达木单抗在治疗过程中易诱发大鼠复发结核,结核发生率高达60%,而模型组和SE-DR亲和肽组未观察到结核复发,提示SE-DR亲和肽在治疗类风湿的同时,不会增加结核感染的发生率,预期可以安全应用于结核阳性的RA患者。具体见实施例7。
实施例7
大鼠潜伏感染模型建立:采用5×10
3CFU Erdman株(CFU指集落形成单位)经静脉感染雌性Lewis大鼠,感染后4周检测大鼠肺、脾组织结核分枝杆菌(MTB)量可达1×10
4以上,此时在饮用水中加入抗结核药物异烟肼(0.1g/L)和利福平(15g/L)持续治疗4周,停药2周,此时大鼠肺、脾内MTB培养阴性,造成结核潜伏感染模型。
采用上述结核潜伏感染的雌性Lewis大鼠,建立结核潜伏感染的CIA模型。方法:使用牛CⅡ同IFA配制的乳剂(含100μg胶原),于结核潜伏感染的大鼠的尾根部皮内注射进行免疫,7天后同法加强免疫1次,此后每日检查大鼠四爪形态变化,炎症评分为1分时随机入组给予治疗。发病大鼠随机分为模型组,SE-DR亲和肽组(分别给予FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37,剂量0.2mmol)和阿达木单抗1mg/kg组,每组10只。另设8只未造模的潜伏感染大鼠作为空白组,模型组和空白组给予PBS,各组大鼠均按5ml/kg的体积尾静脉注射给予相应药物,隔天给药一次,连续给药21天,于给药后第22天处死大鼠。给药期间评价各组大鼠四爪的炎症评分,并在处死时检测大鼠肺、脾MTB培养情况。
大鼠造模后,与空白组相比,成模大鼠的炎症评分明显升高,为了反映药物对该指标整 个病程的影响,并对药物的整体作用效果进行评价,主要对炎症评分曲线下面积(AUC)进行统计分析。结果显示,与模型组相比,SE-DR亲和肽组和阿达木单抗给药组均能够显著降低发病大鼠的炎症评分(P<0.01)。处死时,空白组、模型组和SE-DR亲和肽组大鼠的肺、脾内结核分枝杆菌培养均为阴性,但是阿达木单抗组6/10的大鼠肺、脾内结核分枝杆菌培养阳性。此外,阿达木单抗组给药期间动物有明显的竖毛、活动减少等毒性症状,而SE-DR亲和肽给药后大鼠状态良好。上述结果提示,SE-DR亲和肽不增加大鼠结核复发的风险,安全性优于阿达木单抗。结果见表8。
表8:SE-DR亲和肽对结核潜伏感染的CIA大鼠治疗作用及结核复发情况观察
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
2.3 SE-DR亲和肽无诱发乙肝(HBV)感染的风险
为了进一步对比SE-DR亲和肽与生物制剂在诱发感染方面的差异,本研究采用慢性乙肝感染的CIA小鼠模型,对比阿巴西普和SE-DR亲和肽在治疗小鼠关节炎过程中,小鼠乙肝的激活情况。
结果表明,阿巴西普和SE-DR亲和肽均对CIA小鼠具有良好的治疗作用,但是阿巴西普在治疗过程中易出现乙肝的激活,表现为50%的小鼠出现ALT水平升高至>100IU/ml,而SE-DR亲和肽组、模型组及空白组的ALT无明显差异,提示SE-DR亲和肽在治疗类风湿的同时,不会增加乙肝的激活率,预期可以安全应用于合并乙型肝炎的RA患者。具体见实施例8。
实施例8
小鼠慢性乙肝感染模型建立:采用DBA/1小鼠,将Raav8-1.3HBV(1×10
12vg/mL)病毒经尾静脉注射至小鼠体内,建立HBV慢性乙肝模型。
采用上述结慢性乙肝感染的DBA/1小鼠,建立慢性乙肝感染的CIA模型。方法:于小鼠 尾静脉注射Raav8-1.3HBV(1×10
12vg/mL)病毒后第2天,使用牛CⅡ同CFA配制的乳剂(含100μg胶原)对小鼠尾根部皮内注射进行免疫,21天后采用牛CⅡ同IFA配制的乳剂(含100μg胶原)腹腔注射加强免疫1次。此后每日检查小鼠四爪形态变化,炎症评分为1分时随机入组给予治疗。发病小鼠随机分为模型组,SE-DR亲和肽组(分别给予FNS007(APL2)、APL7、APL9、APL20、ALP21、ALP37,剂量50μmol)和阿巴西普组(剂量100μg),每组12只。另设10只未造模的潜伏乙肝感染小鼠作为空白组,模型组和空白组给予PBS,各组小鼠均按10ml/kg的体积尾静脉注射给予相应药物,隔天给药一次,连续给药27天,于给药后第28天处死小鼠。给药期间评价各组小鼠四爪的炎症评分;于加强免疫当天(尾静脉注射病毒后22天)及最终处死取材时检测小鼠病毒血清学及肝功血清学指标。
小鼠造模后,与空白组相比,成模小鼠的炎症评分明显升高,为了反映药物对该指标整个病程的影响,并对药物的整体作用效果进行评价,主要对炎症评分曲线下面积(AUC)进行统计分析。结果显示,与模型组相比,SE-DR亲和肽组和阿巴西普组均能够显著降低发病小鼠的炎症评分(P<0.01)。
乙肝病毒评估方面,在小鼠加强免疫时,即尾静脉注射病毒后22天,全部小鼠均能检测到HBsAg、HBeAg及HBV DNA阳性,且ALT及AST平均水平分别为45及65U/L。提示已制备成功乙肝慢性感染模型。处死时,空白组、模型组和SE-DR亲和肽组小鼠的肝功血清学指标与加强免疫时水平相当,但是阿巴西普组的肝功血清学指标明显升高,50%的小鼠出现ALT水平升高至>100IU/ml,提示小鼠乙肝激活风险大大升高。此外,阿巴西普给药期间动物有明显的竖毛、活动减少等毒性症状,而SE-DR亲和肽给药后小鼠状态良好。上述结果提示,SE-DR亲和肽不增加小鼠乙肝激活的风险,安全性优于阿巴西普。最终取材时各指标结果见表9。
表9:SE-DR亲和肽对慢性乙肝感染的CIA小鼠治疗作用及乙肝激活情况观察
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
3.联合用药方案用于RA对象的实验结果
抗原特异性疗法与非抗原特异性疗法联合用药为未来的类风湿治疗方向。一方面非抗原特异性疗法可以协助抗原特异性疗法尽快控制临床症状(如炎症等),另一方面最终通过抗原特异性药物的免疫调节作用,重建免疫耐受,彻底控制疾病。
为了验证该理论,本研究采用CIA模型,考察SE-DR亲和肽(抗原特异性治疗药物)与阿达木单抗/阿巴西普/甲氨蝶呤(非抗原特异性治疗药物)联合应用,比较其与SE-DR亲和肽、阿达木单抗、阿巴西普或甲氨蝶呤单独用药的差异。
结果表明:与SE-DR亲和肽单药组相比,SE-DR亲和肽与阿达木单抗联合用药组、SE-DR亲和肽与阿巴西普联合用药组及SE-DR亲和肽与甲氨蝶呤联合用药组能够快速地控制大鼠的炎症反应。与阿达木单抗、阿巴西普或甲氨蝶呤单独用药相比,联合用药组的疗效更为显著,作用更安全,并且停药后,SE-DR亲和肽组和联合用药组可维持治疗效果,而阿达木单抗、阿巴西普或甲氨蝶呤组停药后炎症评分有升高趋势。提示SE-DR亲和肽与非抗原特异性治疗药物(如阿达木单抗/阿巴西普/甲氨蝶呤)联合使用,可以大大提高临床缓解率,并且在停药后能够持续缓解,实现减停药。具体见实施例9、实施例10和实施例11。
实施例9
采用雌性Lewis大鼠,建立CIA模型。方法:使用牛CⅡ同IFA配制的乳剂,于大鼠的尾根部皮内注射进行免疫,7天后同法加强免疫1次,此后每日检查大鼠四爪形态变化,炎症评分为1分时随机入组给予治疗。发病大鼠随机分为三组,即:模型组,SE-DR亲和肽组(采用FNS007,0.2mmol),阿达木单抗组(1mg/kg),和联合用药组(FNS007 0.2mmol+阿达木单抗1mg/kg),每组10只。另设8只未造模的雌性Lewis大鼠作为空白组,模型组和空白组给予PBS,各组大鼠均按5mL/kg的体积尾静脉注射给予相应药物,隔天给药一次,连续给药21天,停药后恢复21天。给药期间评价各组大鼠四爪的炎症评分(隔天一次),计算给药期间的炎症评分曲线下面积(AUC)。停药观察期间,继续评估各组大鼠四爪的炎症评分(隔天一次),并于停药后第22天处死大鼠,取踝关节HE染色后,进行组织损伤评价,具体评分标准如下:0分=正常滑膜组织;1分=滑膜增生,炎性细胞浸润;2分=血管翳形成,软骨侵蚀;3分=大部分软骨破坏伴软骨下骨侵蚀;4分=关节完整性丧失,关节强直。
结果表明,在CIA大鼠模型中,与FNS007单药组相比,FNS007与阿达木单抗联合用药组能够快速地控制大鼠的炎症反应,表现为给药前一周,联合用药组的炎症评分较FNS007组更低,炎症评分AUC从14.26降低为11.93,二者有统计学差异(P<0.05);并且整个给药期间联合用药组的炎症评分AUC显著降低,炎症评分AUC从46.17降低为35.88,二者有统计学差异(P<0.05)。此外,与阿达木单抗相比,联合用药组的疗效更为显著(炎症评分AUC从54.51降低为35.88,二者有统计学差异,P<0.05),并且大鼠感染发生率降低、大鼠竖毛、鼻出血等症状明显改善。同时,于给药21天后停药观察21天,停药期可见FNS007组和联合用药组可以维持停药时的治疗效果,而阿达木单抗组停药后疗效难以维持,体现为炎症评分较停药时显著升高(P<0.01)。结果见表10及图1和图2。
表10:FNS007与阿达木单抗联合用药对CIA大鼠的治疗作用
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
实施例10
采用雄性DBA/1小鼠,建立CIA模型。方法:使用牛CⅡ同CFA配制的乳剂(含100μg胶原)对小鼠尾根部皮内注射进行免疫,21天后采用牛CⅡ同IFA配制的乳剂(含100μg胶原)腹腔注射加强免疫1次。此后每日检查小鼠四爪形态变化,炎症评分为1分时随机入组给予治疗。发病小鼠随机分为三组,即:模型组,SE-DR亲和肽组(采用FNS007,0.2mmol),阿巴西普组(5mg/kg),和联合用药组(FNS007 0.2mmol+阿巴西普5mg/kg),每组12只。另设10只未造模的雌性DBA/1小鼠作为空白组,模型组和空白组给予PBS,各组小鼠均按10mL/kg的体积尾静脉注射给予相应药物,隔天给药一次,连续给药28天,停药后恢复28天。给药期间评价各组小鼠四爪的炎症评分(隔天一次),计算给药期间的炎症评分曲线下面积(AUC)。停药观察期间,继续评估各组小鼠四爪的炎症评分(隔天一次),并于停药后第28天处死小鼠,取踝关节HE染色后,进行组织损伤评价,具体评分标准如下:0分=正常滑膜组织;1分=滑膜增生,炎性细胞浸润;2分=血管翳形成,软骨侵蚀;3分=大部分软骨破坏伴软骨下骨侵蚀;4分=关节完整性丧失,关节强直。
结果表明,在CIA小鼠模型中,与FNS007单药组相比,FNS007与阿巴西普联合用药组能够快速地控制小鼠的炎症反应,表现为给药前10天,联合用药组的炎症评分较FNS007组更低,炎症评分AUC从25.64降低为18.11,二者有统计学差异(P<0.05);并且整个给药期间联合用药组的炎症评分AUC显著降低,炎症评分AUC从90.23降低为75.87,二者有统计学差异(P<0.05)。此外,与阿巴西普组相比,联合用药组的疗效更为显著(炎症评分AUC从89.16降低为75.87,二者有统计学差异,P<0.05),并且小鼠感染发生率降低、小鼠竖毛、鼻出血等症状明显改善。同时,于给药28天后停药观察28天,停药期可见FNS007组和联合用药组可以维持停药时的治疗效果,而阿巴西普组停药后疗效难以维持,体现为炎症评分较停药时显著升高(P<0.01)。结果见表11。
表11:FNS007与阿巴西普联合用药对CIA小鼠的治疗作用
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
实施例11
采用雌性Lewis大鼠,建立CIA模型。方法:使用牛CⅡ同IFA配制的乳剂,于大鼠的尾根部皮内注射进行免疫,7天后同法加强免疫1次,此后每日检查大鼠四爪形态变化,炎症评分为1分时随机入组给予治疗。发病大鼠随机分为三组,即:模型组,SE-DR亲和肽组(采用APL20,0.2mmol),甲氨蝶呤组(0.5mg/kg),和联合用药组(APL20 0.2mmol+甲氨蝶呤0.5mg/kg),每组10只。另设8只未造模的雌性Lewis大鼠作为空白组,模型组和空白组给予PBS,各组大鼠均按5mL/kg的体积尾静脉注射给予相应药物,甲氨蝶呤每周给药2次,其他药物隔天给药一次,连续给药21天,停药后恢复21天。给药期间评价各组大鼠四爪的炎症评分(隔天一次),计算给药期间的炎症评分曲线下面积。停药观察期间,继续评估各组大鼠四爪的炎症评分(隔天一次),并于停药后第22天处死大鼠,取踝关节HE染色后,进行组织损伤评价,具体评分标准如下:0分=正常滑膜组织;1分=滑膜增生,炎性细胞浸润;2分=血管翳形成,软骨侵蚀;3分=大部分软骨破坏伴软骨下骨侵蚀;4分=关节完整性丧失,关节强直。
结果表明,在CIA大鼠模型中,与APL20单药组相比,APL20与甲氨蝶呤联合用药组能够快速地控制大鼠的炎症反应,表现为给药前一周,联合用药组的炎症评分较FNS007组更低,炎症评分AUC从14.93降低为12.06,二者有统计学差异(P<0.05);并且整个给药期间联合用药组的炎症评分AUC显著降低,炎症评分AUC从48.69降低为37.59,二者有统计学差异(P<0.05)。此外,与甲氨蝶呤相比,联合用药组的疗效更为显著(炎症评分AUC从67.69降低为37.59,二者有统计学差异,P<0.05),并且大鼠竖毛、鼻出血等症状明显改善,动物死亡率降低。同时,于给药21天后停药观察21天,停药期可见APL20组和联合用药组可以维持停药时的治疗效果,而甲氨蝶呤组停药后疗效难以维持,体现为炎症评分较停药时显著升高(P<0.01)。结果见表12。
表12:APL20与甲氨蝶呤联合用药对CIA大鼠的治疗作用
注:与空白组比较
#P<0.05,
##P<0.01;与模型组比较
*P<0.05,
**P<0.01。
由此可见,SE-DR亲和肽组与非抗原特异性治疗药物(如阿达木单抗/阿巴西普/甲氨蝶呤)联合使用,可以大大提高临床缓解率,并且在停药后能够持续缓解,实现减停药。因此,本发明能够提供一种全新的能够显著提高临床缓解率并且有望实现减停药的药物组合物。
显然,本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样,倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内,则本发明也意图包含这些改动和变型在内。
Claims (16)
- SE-DR亲和肽在制备治疗结核阳性和/或合并乙型肝炎的风湿疾病患者的风湿疾病的药物中的用途,其特征在于:所述SE-DR亲和肽是指与HLA DR分子的共享表位结合的肽段,优选地,所述共享表位为HLA-DR分子的第70-74位氨基酸,均表达QK/RRAA序列,所述SE-DR亲和肽通过竞争性抑制风湿疾病相关的抗原肽与HLA DR分子的结合发挥作用,但不具备广泛的免疫抑制作用,无诱发感染、肿瘤的风险。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽与SE-DR结合的核心序列包含P1至P9对应的核心氨基酸,其中P1位点为含疏水侧链的氨基酸及有类似性质的稀有或非天然氨基酸,P4位点为非极性、极性不带电荷、极性带负电荷的氨基酸及有类似性质的稀有或非天然氨基酸,优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)、Trp(W)、Leu(L)、Ile(I)、Met(M)、Val(V)或Ala(A)中的一种,所述P4位点选自Met(M)、Ala(A)、Val(V)、Ile(I)、Leu(L)、Asp(D)、Glu(E)、Gln(Q)、Ser(S)或Cit中的一种,更优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)或Trp(W)中的一种,所述P4位点选自Met(M)、Leu(L)、Asp(D)、Glu(E)或Cit中的一种。
- 根据权利要求2所述的用途,其特征在于:所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点为含短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,P9位点为含中、短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点选自Ala(A)、Gly(G)、Ser(S)、Thr(T)或Asn(N)中的一种,所述P9位点选自Ala(A)、Gly(G)、Leu(L)或Met(M)中的一种。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:FXGEQGXXGE,或FXGEXAXXGE,其中:X选自P、K、Q、A或G,优选地,X选自A或G,更优选地,所述SE-DR亲和肽是FNS007(FKGEQAGAGE)。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:YXKQXTXXLA,其中:X选自V、A、G、N、L或K,优选地,X选自A或G,更优选地,所述SE-DR亲和肽的氨基酸序列选自PKYVKQNTLKLAT、PGYVKQGTLGLAT、YVKQNTLKLA、YVAQNTLKLA、YAKQATLKLA或YAKQATLALA。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:IWYIXCFXCEXHXXL,其中:X选自A、G、M、T、V、N、Q或S,优选地,X选自A或G,更优选地,所述SE-DR亲和肽的氨基酸序列选自IWYINCFGCETHAML、IWYIQCFGCETHAML、IWYISCFGCETHAML、IWYITCFGCETHAML、IWYINCFACETHAML或IWYINCFVCETHAML。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:RSFXLAXSXXGVG,其中:X选自A、G、T、S或E,优选地,X选自A或G,更优选地,所述SE-DR亲和肽的氨基酸序列选自RSFTLASSETGVG、RSFALASSETGVG、RSFTAASSETGVG、RSFTLDSSETGVG、RSFTLAASETGVG、RSFTLASSATGVG、RSFTLASSEAGVG或RSFTLDSSETGVG。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:SAVXLCitXSXXGVR,其中:X选自A、G、R、S、V或P,优选地,所述SE-DR亲和肽的氨基酸序列为SAVRLCitSSVPGVR、SAVELCitSSVPGVR、SAVDLCitSSVPGVR、SAVGLCitSSVPGVR、SAVRLCitFSVPGVR、SAVRLCitSSVEGVR、SAVRLCitSSVKGVR、SAVRLCitSSVWGVR、SAVRLCitWSVPGVR、SAVRLCitSSVRGVR、SAVRLCitKSVPGVR、SAVALCitSSVPGVR或SAVRLCitRSVPGVR;或者,所述SE-DR亲和肽包含下述氨基酸序列:GVYXTCitXSXXCitLCit,其中:X选自A、G或V,优选地,X选自A或G,更优选地,所述SE-DR亲和肽的氨基酸序列为GVYATCitSSAVCitLCit;或者,所述SE-DR亲和肽包含下述氨基酸序列:QDXNCitXNXXKNS,其中:X选自A、G、F、I、K或L,优选地,X选自A或G,更优选地,所述SE-DR亲和肽的氨基酸序列为QDFTNCitANKLKNS;或者,所述SE-DR亲和肽包含下述氨基酸序列:VVLLVATXGCitXRXXSAYQDK,其中:X选自A、G、E、V或N,优选地,X选自A或G,更优选地,所述 SE-DR亲和肽的氨基酸序列为VVLLVATEGCitVRVNSAYQDK。
- 根据权利要求1所述的用途,其特征在于:所述SE-DR亲和肽的氨基酸序列选自:MGPKGRTVIIEQSWGSPKVTK、MGPKGRTVIIEQSLGSPKVTK、SIDLKDKKYKNIGAKLVQDVANNTNEEA、SIDLKDKKYKNIGAKLVQLVANNTNEEA、QYMCitADQAAGGLR、LTQCitGSVLR、WYNCitCHAAN、VETCitDGQVI或VCitLCitSSVESTCitGRSCitPAPPPACitGLT。
- 根据权利要求1-9中任一项所述的用途,其特征在于:所述风湿疾病选自类风湿关节炎、银屑病、银屑病关节炎、强直性脊柱炎、未分化脊柱关节病、系统性红斑狼疮、系统性硬化或胶原病中的一种或多种。
- 一种治疗风湿疾病的药物组合物,其特征在于:所述药物组合物包含SE-DR亲和肽和非抗原特异性抗风湿疾病药物,以及药学上可接受的载体,所述SE-DR亲和肽是指与HLA DR分子的共享表位结合的肽段,优选地,所述共享表位为HLA-DR分子的第70-74位氨基酸,均表达QK/RRAA序列,所述SE-DR亲和肽通过竞争性抑制风湿疾病相关的抗原肽与HLA DR分子的结合发挥作用,改善患者体内紊乱的免疫微环境,重建免疫平衡,所述药物组合物快速抑制风湿疾病的进程,并且在减停药之后实现持续缓解。
- 根据权利要求11所述的药物组合物,所述SE-DR亲和肽与SE-DR结合的核心序列包含P1至P9对应的核心氨基酸,其中P1位点为含疏水侧链的氨基酸及有类似性质的稀有或非天然氨基酸,优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P1位点选自Tyr(Y)、Phe(F)、Trp(W)、Leu(L)、Ile(I)、Met(M)、Val(V)或Ala(A)中的一种;所述P4位点为非极性、极性不带电荷、极性带负电荷的氨基酸及有类似性质的稀有或非天然氨基酸,优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P4位点选自Met(M)、Ala(A)、Val(V)、Ile(I)、Leu(L)、Asp(D)、Glu(E)、Gln(Q)或Ser(S)中的一种;所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点为含短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,所述P9位点为含中、短侧链的氨基酸及有类似性质的稀有或非天然氨基酸,优选地,所述SE-DR亲和肽与SE-DR结合的核心序列中相应的P6位点选自Ala(A)、Gly(G)、Ser(S)、Thr(T)或Asn(N)中的一种,所述P9位点选自Ala(A)、Gly(G)、Leu(L)或Met(M)中的一种;所述非抗原特异性抗风湿疾病药物选自针对细胞因子靶点、针对B细胞靶点、针对T细胞靶点、针对激酶靶点的药物、传统改善病情的抗风湿药(DMARD)、非甾体抗炎药(NSAID)或激素类药物的一种或多种,优选地,所述非抗原特异性抗风湿疾病药物选自肿瘤坏死因子(TNF)抑制剂、IL-6抑制剂、IL-1抑制剂、IL-17抑制剂、RANKL抑制剂、T细胞共刺激信号抑制剂、CD20抑制剂、CD22抑制剂、BLyS及APRIL抑制剂、BAFF抑制剂、JAK抑制剂、BTK抑制剂、Syk抑制剂、IRAK4抑制剂、p38抑制剂或小分子免疫抑制剂的一种或多种,更优选地,所述非抗原特异性抗风湿疾病药物选自阿达木单抗或其生物类似物、托珠单抗或其生物类似物、阿那白滞素或其生物类似物、托法替尼或其生物类似物、阿巴西普或其生物类似物、利妥昔单抗或其生物类似物、甲氨蝶呤或来氟米特的一种或多种。
- 根据权利要求11或权利要求12所述的药物组合物,其特征在于:所述SE-DR亲和肽包含下述氨基酸序列:FXGEQGXXGE,或FXGEXAXXGE,其中:X选自K、Q、A或G,优选地,X选自A或G,更优选地,所述SE-DR亲和肽是FNS007(FKGEQAGAGE),或者,所述SE-DR亲和肽的氨基酸序列选自:MGPKGRTVIIEQSWGSPKVTK、MGPKGRTVIIEQSLGSPKVTK、SIDLKDKKYKNIGAKLVQDVANNTNEEA、SIDLKDKKYKNIGAKLVQLVANNTNEEA、QYMCitADQAAGGLR、LTQCitGSVLR、WYNCitCHAAN、VETCitDGQVI或VCitLCitSSVESTCitGRSCitPAPPPACitGLT。
- 根据权利要求11-13中任一项所述的药物组合物,其特征在于:所述药学上可接受的载体包括粘合剂、表面活性剂、增溶剂、稳定剂、润滑剂、湿润剂和/或稀释剂,所述药物组合物的剂型为胶囊剂、片剂、丸剂、液剂、散剂、颗粒剂、细粒剂、膜衣剂、沉淀剂、含片剂、舌下剂、咀嚼剂、颊剂、糊剂、糖浆剂、悬浮剂、酏剂、乳剂、涂布剂、软膏剂、硬膏剂、敷剂、经皮吸收型制剂、洗剂、吸入剂、气雾剂、注射剂或栓剂。
- 根据权利要求11-14中任一项所述的药物组合物,其特征在于:所述药物组合物适于通过多种途径施用,包括口服、静脉内、皮下、肌内、脑内、鼻内、肺部、动脉内、关节内、皮内、玻璃体内、骨内输注、腹膜内、鞘内或透皮施用。
- 根据权利要求11-15中任一项所述的药物组合物,其特征在于:所述风湿疾病选自类风湿关节炎、青少年特发性关节炎、银屑病、银屑病关节炎、强直性脊柱炎、未分化脊柱关节病、系统性红斑狼疮、系统性硬化、胶原病、克罗恩病、溃疡性结肠炎的一种或多种,优选地,所述风湿疾病的患者是结核阳性的或结核阴性的;所述风湿疾病的患者是合并乙型肝炎的或者不合并乙型肝炎的。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1386754A (zh) * | 2002-06-27 | 2002-12-25 | 北京大学人民医院 | 非t细胞结合肽及其用途 |
CN1868537A (zh) * | 2006-02-09 | 2006-11-29 | 北京大学人民医院 | Ⅱ型胶原变构肽对类风湿关节炎的治疗作用 |
CN1872873A (zh) * | 2006-06-14 | 2006-12-06 | 北京大学人民医院 | T细胞抑制肽对类风湿关节炎的治疗作用 |
CN1935133A (zh) * | 2005-09-19 | 2007-03-28 | 北京大学 | 人白细胞抗原dr4亚型蛋白的拮抗剂 |
CN103163302A (zh) * | 2011-12-10 | 2013-06-19 | 石家庄恩泽药品技术开发有限公司 | 一种采用定向交叉偶联技术制备的短肽抗体试剂盒 |
CN111868073A (zh) * | 2019-05-31 | 2020-10-30 | 广州市雷德生物科技有限公司 | 一种类风湿关节炎相关的特异性多肽及其应用 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CU23504A1 (es) * | 2004-09-24 | 2010-04-13 | Ct Ingenieria Genetica Biotech | Péptidos y derivados tipo apl de la hsp60 y composiciones farmacéuticas |
CN101244257A (zh) * | 2007-12-19 | 2008-08-20 | 北京大学人民医院 | Ⅱ型胶原变构肽鼻粘膜给药治疗类风湿关节炎的应用 |
CU23701A1 (es) * | 2008-12-29 | 2011-09-21 | Ct Ingenieria Genetica Biotech | Método de tratamiento de enfermedades inflamatorias intestinales y diabetes tipo i |
CN102499980A (zh) * | 2011-12-27 | 2012-06-20 | 中国药科大学 | 多肽在制备治疗或预防类风湿性关节炎药物中的应用 |
EP2968435B1 (en) * | 2013-03-12 | 2019-01-30 | Tel HaShomer Medical Research Infrastructure and Services Ltd. | Synthetic peptides for the treatment of autoimmune diseases |
CA2899577C (en) * | 2013-04-03 | 2023-10-17 | Ibc Pharmaceuticals, Inc. | Combination therapy for inducing immune response to disease |
CN105816852B (zh) * | 2015-01-04 | 2019-11-22 | 河北菲尼斯生物技术有限公司 | 含非t细胞结合肽和乳酸羟基乙酸嵌段共聚物的药物组合物、其制备方法和用途 |
KR102276941B1 (ko) * | 2018-07-03 | 2021-07-14 | 서울대학교산학협력단 | 류마티스 관절염 치료용 펩티드 및 그의 용도 |
-
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- 2021-07-30 CN CN202110873116.6A patent/CN115671253B/zh active Active
- 2021-07-30 CN CN202410144868.2A patent/CN117883547A/zh active Pending
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- 2022-09-07 WO PCT/CN2022/117495 patent/WO2023006125A1/zh active Application Filing
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1386754A (zh) * | 2002-06-27 | 2002-12-25 | 北京大学人民医院 | 非t细胞结合肽及其用途 |
CN1935133A (zh) * | 2005-09-19 | 2007-03-28 | 北京大学 | 人白细胞抗原dr4亚型蛋白的拮抗剂 |
CN1868537A (zh) * | 2006-02-09 | 2006-11-29 | 北京大学人民医院 | Ⅱ型胶原变构肽对类风湿关节炎的治疗作用 |
CN1872873A (zh) * | 2006-06-14 | 2006-12-06 | 北京大学人民医院 | T细胞抑制肽对类风湿关节炎的治疗作用 |
CN103163302A (zh) * | 2011-12-10 | 2013-06-19 | 石家庄恩泽药品技术开发有限公司 | 一种采用定向交叉偶联技术制备的短肽抗体试剂盒 |
CN111868073A (zh) * | 2019-05-31 | 2020-10-30 | 广州市雷德生物科技有限公司 | 一种类风湿关节炎相关的特异性多肽及其应用 |
Non-Patent Citations (3)
Title |
---|
CHICZ, R. M. ET AL.: "Specificity and Promiscuity among Naturally Processed Peptides Bound to HLA-DR Alleles", JOURNAL OF EXPERIMENTAL MEDICINE, vol. 178, 31 July 1993 (1993-07-31), pages 27 - 47, XP002069888, DOI: 10.1084/jem.178.1.27 * |
LI ZHANGUO, HAN LEI , JIA RULIN, LI JING: "Impact of rheumatoid arthritis -associated HLA -DRβ 1subtypes on protein kinase A signaling", CHINESE MEDICAL JOURNAL, vol. 116, no. 5, 31 December 2003 (2003-12-31), pages 712 - 716, XP093028654 * |
YAO Z.-Q., LI R., LI Z.-G.: "A triple altered collagen II peptide with consecutive substitutions of TCR contacting residues inhibits collagen-induced arthritis", ANNALS OF THE RHEUMATIC DISEASES, BRITISH MEDICAL ASSOCIATION, GB, vol. 66, no. 3, 1 March 2007 (2007-03-01), GB , pages 423 - 424, XP093028643, ISSN: 0003-4967, DOI: 10.1136/ard.2006.057927 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117085118A (zh) * | 2023-08-22 | 2023-11-21 | 北京大学人民医院 | 一种瓜氨酸化ii型胶原多肽疫苗及其应用 |
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