CN115708872A - Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor - Google Patents
Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor Download PDFInfo
- Publication number
- CN115708872A CN115708872A CN202211374146.3A CN202211374146A CN115708872A CN 115708872 A CN115708872 A CN 115708872A CN 202211374146 A CN202211374146 A CN 202211374146A CN 115708872 A CN115708872 A CN 115708872A
- Authority
- CN
- China
- Prior art keywords
- antibody
- angiopoietin
- protein
- bone
- inflammatory factor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 29
- 230000002757 inflammatory effect Effects 0.000 title claims abstract description 23
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 19
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 210000000988 bone and bone Anatomy 0.000 claims abstract description 25
- 108090001005 Interleukin-6 Proteins 0.000 claims abstract description 24
- 230000006378 damage Effects 0.000 claims abstract description 17
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 13
- 201000010099 disease Diseases 0.000 claims abstract description 12
- 206010003246 arthritis Diseases 0.000 claims abstract description 4
- 206010031149 Osteitis Diseases 0.000 claims abstract 2
- 208000018937 joint inflammation Diseases 0.000 claims abstract 2
- 102000004889 Interleukin-6 Human genes 0.000 claims description 23
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 23
- 229940100601 interleukin-6 Drugs 0.000 claims description 17
- 102000045205 Angiopoietin-Like Protein 4 Human genes 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 11
- 208000027866 inflammatory disease Diseases 0.000 claims description 6
- 239000002671 adjuvant Substances 0.000 claims description 5
- 230000001684 chronic effect Effects 0.000 claims description 5
- 239000012634 fragment Substances 0.000 claims description 4
- 101710085845 Angiopoietin-related protein 4 Proteins 0.000 claims description 3
- 206010027476 Metastases Diseases 0.000 claims description 3
- 206010028980 Neoplasm Diseases 0.000 claims description 3
- 230000009401 metastasis Effects 0.000 claims description 3
- 230000017074 necrotic cell death Effects 0.000 claims description 3
- 201000004595 synovitis Diseases 0.000 claims description 3
- 206010005963 Bone formation increased Diseases 0.000 claims description 2
- 206010006811 Bursitis Diseases 0.000 claims description 2
- 206010034464 Periarthritis Diseases 0.000 claims description 2
- 208000006735 Periostitis Diseases 0.000 claims description 2
- 239000003963 antioxidant agent Substances 0.000 claims description 2
- 230000003078 antioxidant effect Effects 0.000 claims description 2
- 239000000022 bacteriostatic agent Substances 0.000 claims description 2
- 239000011248 coating agent Substances 0.000 claims description 2
- 238000000576 coating method Methods 0.000 claims description 2
- 239000003086 colorant Substances 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000003995 emulsifying agent Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 230000003204 osmotic effect Effects 0.000 claims description 2
- 201000008482 osteoarthritis Diseases 0.000 claims description 2
- -1 pH regulator Substances 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 239000004094 surface-active agent Substances 0.000 claims description 2
- 239000000080 wetting agent Substances 0.000 claims description 2
- 239000000872 buffer Substances 0.000 claims 1
- 239000007884 disintegrant Substances 0.000 claims 1
- 208000009386 Experimental Arthritis Diseases 0.000 abstract description 23
- 241000699666 Mus <mouse, genus> Species 0.000 abstract description 16
- 238000010171 animal model Methods 0.000 abstract description 12
- 210000000544 articulatio talocruralis Anatomy 0.000 abstract description 10
- 210000001503 joint Anatomy 0.000 abstract description 8
- 230000008595 infiltration Effects 0.000 abstract description 7
- 238000001764 infiltration Methods 0.000 abstract description 7
- 210000004969 inflammatory cell Anatomy 0.000 abstract description 7
- 229940079593 drug Drugs 0.000 abstract description 6
- 206010020718 hyperplasia Diseases 0.000 abstract description 5
- 230000001225 therapeutic effect Effects 0.000 abstract description 5
- 210000001258 synovial membrane Anatomy 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 17
- 101150054149 ANGPTL4 gene Proteins 0.000 description 11
- 108700042530 Angiopoietin-Like Protein 4 Proteins 0.000 description 11
- 230000000694 effects Effects 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 9
- 210000002683 foot Anatomy 0.000 description 7
- 230000008859 change Effects 0.000 description 6
- 238000010603 microCT Methods 0.000 description 6
- 238000010186 staining Methods 0.000 description 6
- 206010023232 Joint swelling Diseases 0.000 description 5
- 230000008961 swelling Effects 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 208000017170 Lipid metabolism disease Diseases 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 210000000845 cartilage Anatomy 0.000 description 3
- 230000003285 pharmacodynamic effect Effects 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 208000010392 Bone Fractures Diseases 0.000 description 2
- 102000000503 Collagen Type II Human genes 0.000 description 2
- 108010041390 Collagen Type II Proteins 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 206010028851 Necrosis Diseases 0.000 description 2
- 125000003275 alpha amino acid group Chemical group 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 210000003423 ankle Anatomy 0.000 description 2
- 230000006399 behavior Effects 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 229940030156 cell vaccine Drugs 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000000770 proinflammatory effect Effects 0.000 description 2
- 210000004989 spleen cell Anatomy 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010017076 Fracture Diseases 0.000 description 1
- 241001111421 Pannus Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000002591 computed tomography Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000009707 neogenesis Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000005222 synovial tissue Anatomy 0.000 description 1
- 210000002437 synoviocyte Anatomy 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- 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]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physical Education & Sports Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Rheumatology (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Pain & Pain Management (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses an application of an anti-angiopoietin-like protein 4 antibody in preparing a medicament for inhibiting the expression of an inflammatory factor. According to the invention, by constructing a mouse animal model of collagen-induced arthritis (CIA), simulating the disease state of RA and applying different doses of anti-ANGPTL 4 antibody to perform therapeutic intervention on the CIA model, the anti-ANGPTL 4 antibody treatment can obviously reduce the expression of a cytokine IL-6, can effectively improve the bone destruction of RA on ankle joints, and can relieve the synovial membrane hyperplasia of local joints and infiltration of inflammatory cells, so that the anti-ANGPTL 4 antibody can be applied to the preparation of medicines for treating the diseases of anti-bone and joint inflammation.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and relates to application of an anti-angiopoietin-like protein 4 antibody in preparation of a medicament for inhibiting expression of an inflammatory factor.
Background
Rheumatoid Arthritis (RA) is a chronic systemic disease with inflammatory synovitis as the main cause, which is characterized by polyarticular, symmetrical and aggressive arthritis of hand and foot small joints, often accompanied by the involvement of extra-articular organs and positive serum rheumatoid factors, and can cause joint deformity and function loss. The pathology of RA arthritis is mainly synovial lining cell hyperplasia, interstitial massive inflammatory cell infiltration, microvascular neogenesis, pannus formation, cartilage and bone tissue destruction, and the like.
Currently, biological compounds, such as antibodies, targeting tumor necrosis factor alpha (TNF- α), B cells, or T cells have been used to treat RA, but many patients fail to respond to these therapies. For example, CN104995210A discloses an anti-GM-CSF antibody for the treatment of rheumatoid arthritis. CN110585428A discloses the application of B cell vaccine in preparing drugs for treating rheumatoid arthritis, wherein the B cell vaccine is prepared from Non-B10B cells obtained by sorting spleen cells of collagen-induced arthritis mice; the sorting method comprises the following steps: the spleen cells are firstly subjected to surface staining by antibodies CD19-FITC, CD5-APC and CD1d-PE, then flow cytometry sorting is carried out, all B cells except B10 cells (CD 19+ CD5+ CD1 dhi) are collected, and Non-B10B cells are obtained; the preparation method comprises the following steps: culturing Non-B10B cells under stimulation in the presence of functional antibodies anti-CD40, cpG, type II Collagen (CII) and BAFF, and irradiating with cobalt source to obtain vaccine.
ANGPTL4 contains an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain, and has 3 active forms in vivo, which are full-length angiopoietin-like protein 4 (flANGPTL 4), N-terminal (nANGPTL 4) and C-terminal (cANGPTL 4) fragments, respectively. Antibodies capable of neutralizing at least one active form of ANGPTL4 are widely used in the treatment of lipid metabolism disorders, e.g., CN101128485 discloses a monoclonal antibody against ANGPTL4, and also provides the use of the monoclonal antibody in the preparation of a medicament for treating a lipid metabolism disorder in a patient.
In conclusion, the novel medicine for treating the rheumatoid arthritis is developed, a new thought and a new means are provided for treating the rheumatoid arthritis, and the medicine has important significance for the field of treating the rheumatoid arthritis.
Disclosure of Invention
The invention provides the application of an anti-angiopoietin-like protein 4 antibody in preparing a medicament for inhibiting the expression of inflammatory factors aiming at the problem that most of rheumatoid arthritis medicaments in clinic have single action effect and the requirement on novel rheumatoid arthritis treatment medicaments, and finds that the anti-angiopoietin-like protein 4 can play an anti-inflammatory role by reducing proinflammatory cytokine interleukin-6 (IL-6), has the function of inhibiting bone destruction and can be applied to preparing medicaments for treating bone and joint inflammatory diseases.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides the use of an anti-angiopoietin-like protein 4 antibody in the preparation of a medicament for inhibiting the expression of an inflammatory factor.
In the invention, pharmacodynamic verification of a medicament is carried out by a classical collagen-induced arthritis (CIA) animal model in the RA research field, in the experimental process, the alleviation effect of the anti-ANGPTL 4 antibody on joint swelling is evaluated by measuring body weight and paw swelling, and serum detection of the animal model shows that the anti-ANGPTL 4 antibody treatment can obviously reduce the expression of inflammatory factors.
Preferably, the inflammatory factor comprises interleukin-6 (IL-6).
In the invention, the anti-angiopoietin-like protein 4 antibody refers to an antibody which is combined with any one of a flANGPTL4 fragment, a nANGPTL4 fragment or a cANGPTL4 of full-length angiopoietin-like protein 4 in a targeted mode, and the clinical application range of the existing anti-ANGPTL 4 antibody is mainly drugs for treating lipid metabolism diseases.
It is understood that anti-ANGPTL 4 antibodies disclosed in the art are suitable for use in the present invention, and in particular, the anti-ANGPTL 4 antibodies can be selected from humanized monoclonal antibodies, see US10160803B2 for specific amino acid sequences.
Preferably, the medicament further comprises an adjuvant.
Preferably, the adjuvant comprises any one or a combination of at least two of a pharmaceutically acceptable carrier, a wetting agent, a disintegrating agent, an emulsifier, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent or a buffering agent.
In a second aspect, the invention provides the use of an anti-angiopoietin-like protein 4 antibody in the preparation of a medicament for the treatment of chronic bone and joint inflammatory diseases.
The invention utilizes a collagen-induced arthritis (CIA) animal model to perform pharmacodynamic verification of a medicament for resisting the ANGPTL4 antibody, and Micro-CT scanning data and histological staining results show that the ANGPTL4 antibody can effectively improve the bone destruction of ankle joints of RA and relieve the synovial hyperplasia of local joints and infiltration of inflammatory cells.
Preferably, the chronic bone and joint inflammatory disease includes any one or a combination of at least two of rheumatoid arthritis, osteoarthritis, hyperosteogeny, scapulohumeral periarthritis, periostitis, bursitis or synovitis.
In a third aspect, the invention provides the use of an anti-angiopoietin-like protein 4 antibody in the preparation of a medicament for the treatment of a bone destruction disease.
Preferably, the bone destruction disease comprises any one of femoral head necrosis, refractory fracture or tumor bone metastasis or a combination of at least two of them.
According to the invention, a mouse animal model of collagen-induced arthritis (CIA) is constructed, and the anti-ANGPTL 4 antibody with different doses is applied to perform therapeutic intervention on the CIA model, so that the result shows that the anti-ANGPTL 4 antibody can relieve the progress of diseases, and the result of imaging and histology shows that the ankle joint of a mouse treated by the anti-ANGPTL 4 antibody obviously improves the bone destruction condition, so that the collagen-induced arthritis (CIA) can be used for developing and treating related diseases such as femoral head necrosis, bone fracture difficult to heal, bone destruction caused by tumor bone metastasis and the like.
In a fourth aspect, the invention provides a non-therapeutic use of an anti-angiopoietin-like protein 4 antibody in the preparation of an inhibitor of inflammatory factor expression.
In the present invention, the inhibitor refers to an agent capable of inhibiting the expression of an inflammatory factor.
In the invention, the anti-angiopoietin-like protein 4 antibody is found to be capable of regulating the expression level of proinflammatory cytokine interleukin-6 (IL-6) in vivo, and can be used as an inhibitor to be applied to the basic research of interleukin-6 (IL-6) related behaviors.
In a fifth aspect, the present invention provides a method of inhibiting the expression of an inflammatory factor for the purpose of non-disease diagnosis and/or treatment, the method comprising:
the anti-angiopoietin-like protein 4 antibody is used for preparing the antibody for inhibiting the expression of inflammatory factors, wherein the inflammatory factors comprise interleukin-6.
In the invention, the anti-angiopoietin-like protein 4 antibody can be used as an inhibitor to be applied to basic research of interleukin-6 (IL-6) related behaviors, such as constructing an interleukin-6 (IL-6) low-expression animal model.
Compared with the prior art, the invention has the following beneficial effects:
the invention discovers for the first time that the anti-ANGPTL 4 antibody can inhibit the expression of inflammatory factor interleukin-6 (IL-6), can relieve the synovial hyperplasia of local joints and the infiltration of inflammatory cells, has the function of inhibiting bone destruction, can be applied to the preparation of medicines for treating bone and joint inflammatory diseases, overcomes the defect of single effect of most RA medicines clinically at present, provides a new thought and a new means for treating rheumatoid arthritis, and has important significance for the field of treating the rheumatoid arthritis.
Drawings
FIG. 1 is a graph of the change in body weight of mice;
FIG. 2 is a graph of the results of clinical scores for joint swelling in mice;
FIG. 3 is a graph showing the expression level of ANGPTL4 in mouse serum;
FIG. 4 is a graph showing the expression level of IL-6 in mouse serum;
FIG. 5 is a three-dimensional view of mouse ankle joint Micro-CT;
FIG. 6 is a histological staining chart of pathological sections of mouse ankle joints.
Detailed Description
To further illustrate the technical means adopted by the present invention and the effects thereof, the present invention is further described below with reference to the embodiments and the accompanying drawings. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and that no limitation of the invention is intended.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or apparatus used are conventional products commercially available from normal sources, not indicated by the manufacturer.
The invention specifically carries out pharmacodynamic verification of the medicament through a classical CIA animal model in the RA research field. In the experimental process, the alleviation effect of the anti-ANGPTL 4 antibody on joint swelling and the toxicity effect under the treatment dosage are evaluated by measuring the body weight and paw swelling, and the expression of the cytokine IL-6 is analyzed by serological detection collected from an animal model; the effect of anti-ANGPTL 4 antibody treatment on bone destruction of the ankle joint of RA was analyzed by Micro-CT scan data and histological staining results.
Example 1
This example was performed for model construction and therapeutic intervention with anti-ANGPTL 4 antibodies.
32 SPF-grade 8-week-old male DBA/1 immune normal mice are purchased, the mice are raised in an SPF-grade animal room, the mice are randomly grouped according to the average weight, 8 mice are divided into a blank control group (CON), a model group (CIA), a low-dose treatment group (50 mg/kg) and a high-dose treatment group (200 mg/kg), an immune program inducer containing 2mg/mL bovine type II collagen and 4mg/mL complete Freund's adjuvant is prepared, after full emulsification, 100 mu L of the immune program inducer is subcutaneously injected at the tail of the mice in the other groups except the blank control group, the CIA model is constructed, after 3 weeks after modeling, anti-ANGPTL 4 antibody (amino acid sequence is shown in US10160803B 2) is injected in an intraperitoneal mode according to the above dosage, 2 times of weekly administration is carried out, the model group is injected as a control until the end point of the model group, the whole experiment is maintained for 7 weeks, the weight and the foot paw swelling is measured, the result is shown in figure 1, after modeling, after 3 weeks, the mice are subjected to the normal mouse injection, the mice are subjected to the normal mouse administration of PBS, the normal mouse treatment, the mice in the normal mouse growth rate is obviously reduced in the third week, and the normal mouse treatment of the mice after the normal mouse growth of the normal mice is shown in the normal treatment. Joint swelling clinical scores (specific method references: n. Choudhary, l.k.bhatt, and k.s.prabhavalvar, experimental animal models for rheumatoid arthritis. Immunopharmacology and immunovirology 40 (2018) 193-200.), results are shown in fig. 2, indicating that anti-ANGPTL 4 antibody treatment can alleviate clinical symptoms of paw swelling in CIA animal models.
Example 2
In this example, the expression level of ANGPTL4 and the change in the expression of inflammatory factors were measured.
After 7 weeks of the model building in example 1, the mouse was anesthetized, the heart was bled, and the serum was obtained by standing and separating, and the change in the expression level of ANGPTL4 and the change in the content of cytokine IL-6 in the serum were detected by using an ELISA detection kit, and the result of the change in the expression level of ANGPTL4 is shown in fig. 3, and as compared with the CIA group, the expression levels of ANGPTL4 in the mouse sera of the low dose treatment group (50 mg/kg) and the high dose treatment group (200 mg/kg) were significantly reduced, indicating that the ANGPTL4 in the serum could be effectively neutralized. The results of the content change of the cytokine IL-6 are shown in FIG. 4, and the results show that the expression level of the inflammatory factor IL-6 in the serum of the mice is reduced after the treatment with 200mg/kg of anti-ANGPTL 4 antibody compared with the CIA model group.
Example 3
This example was subjected to Micro-CT analysis.
The mice were euthanized 7 weeks after the model of example 1, ankle tissue was dissected and isolated, anterior and posterior tibial muscles were removed, and the tibia was interrupted and cut off, leaving the intact paw and ankle of the mice and fixed with 10% formalin. Scanning the ankle joints of the mice by using Micro-CT (SKYSCAN 1176, bruker, belgium), reconstructing scanning data, and observing the damage degree of each group of joints, wherein the result is shown in figure 5, and the surfaces of the joints of the CON group are smooth and complete; after 50mg/kg and 200mg/kg of anti-ANGPTL 4 antibody treatment, the destruction condition of the phalangeal joints is improved, which indicates that the anti-ANGPTL 4 antibody has the treatment effect of inhibiting inflammatory bone destruction, and the treatment effect is dose-dependent.
Example 4
This example was performed for histological staining analysis of the local part of the joint.
Sequentially decalcifying, dehydrating and conventional paraffin embedding bone tissues subjected to CT scanning in example 3, and then making tissue sections and performing HE staining, wherein the results are shown in figure 6, the surfaces of ankle joint cartilages of animals in CON group are smooth, joint cavities are complete, and joint gaps are obvious; a large amount of inflammatory cell infiltration can be seen at the ankle joint of the CIA modeling animal group, the joint gap is narrowed, the cartilage surface is damaged, and the synovial tissue is proliferated; in the anti-ANGPTL 4 antibody treatment group, the joint cavity destruction degree and the inflammatory cell infiltration degree are relieved, and the bone destruction degree is relieved.
In conclusion, the disease state of RA is simulated by constructing a mouse animal model of collagen-induced arthritis (CIA), the CIA model is subjected to therapeutic intervention by using anti-ANGPTL 4 antibodies with different doses, the alleviation effect of the anti-ANGPTL 4 antibodies on joint swelling and the toxic effect under the therapeutic dose are evaluated by measuring the weight and paw swelling, and serum detection of the animal model shows that the expression of a cytokine IL-6 can be obviously reduced by the anti-ANGPTL 4 antibody treatment; micro-CT scanning data and histological staining results show that the anti-ANGPTL 4 antibody can effectively improve the bone destruction of the ankle joint of RA, relieve the synovial hyperplasia of the local joints and the infiltration of inflammatory cells, and prove that the anti-ANGPTL 4 antibody can be applied to preparing medicaments for treating bone and joint inflammatory diseases.
The applicant states that the present invention is illustrated in detail by the above examples, but the present invention is not limited to the above detailed methods, i.e. it is not meant that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (10)
1. Application of anti-angiopoietin-like protein 4 antibody in preparing medicine for inhibiting inflammatory factor expression.
2. The use of claim 1, wherein the inflammatory factor comprises interleukin-6.
3. The use according to claim 1 wherein the anti-angiopoietin-like protein 4 antibody targets any of the flANGPTL4, nANGPTL4, or cANGPTL4 fragments that bind angiopoietin-like protein 4.
4. The use of claim 1, wherein the medicament further comprises an adjuvant.
5. The use according to claim 4, wherein the adjuvant comprises any one or a combination of at least two of a pharmaceutically acceptable carrier, wetting agent, disintegrant, emulsifier, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH regulator, antioxidant, bacteriostatic agent, or buffer.
6. Application of anti-angiopoietin-like protein 4 antibody in preparing medicine for treating chronic bone and joint inflammation diseases.
7. The use according to claim 6, wherein the chronic bone and joint inflammatory disease comprises any one or a combination of at least two of rheumatoid arthritis, osteoarthritis, hyperosteogeny, scapulohumeral periarthritis, periostitis, bursitis or synovitis.
8. Use of an anti-angiopoietin-like protein 4 antibody in the preparation of a medicament for the treatment of a bone-destroying disease.
9. The use of claim 8, wherein the bone destruction disease comprises any one of femoral head necrosis, refractory fracture, or tumor bone metastasis or a combination of at least two of the foregoing.
10. A method of inhibiting the expression of an inflammatory factor for purposes other than disease diagnosis and/or treatment, the method comprising:
inhibiting inflammatory factor expression using anti-angiopoietin-like protein 4 antibodies;
the inflammatory factor comprises interleukin-6.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211374146.3A CN115708872A (en) | 2022-11-03 | 2022-11-03 | Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor |
| PCT/CN2022/132616 WO2024092888A1 (en) | 2022-11-03 | 2022-11-17 | Use of anti-angiopoietin-like protein 4 antibody in preparation of drug for inhibiting expression of inflammatory factor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211374146.3A CN115708872A (en) | 2022-11-03 | 2022-11-03 | Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115708872A true CN115708872A (en) | 2023-02-24 |
Family
ID=85232126
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211374146.3A Pending CN115708872A (en) | 2022-11-03 | 2022-11-03 | Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115708872A (en) |
| WO (1) | WO2024092888A1 (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101387781B1 (en) * | 2005-01-07 | 2014-04-21 | 렉시컨 파마슈티컬스 인코퍼레이티드 | Monoclonal antibodies against angiopoietin-like protein 4 (angptl4) |
| GB201301313D0 (en) * | 2013-01-25 | 2013-03-06 | Univ Singapore | Respiratory Tract Infections |
-
2022
- 2022-11-03 CN CN202211374146.3A patent/CN115708872A/en active Pending
- 2022-11-17 WO PCT/CN2022/132616 patent/WO2024092888A1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| WO2024092888A1 (en) | 2024-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2007287510B2 (en) | Treatment of cartilage disorders with FGF-18 | |
| Zhao et al. | Spinal interleukin-33 and its receptor ST2 contribute to bone cancer-induced pain in mice | |
| Loffredo et al. | Targeted delivery to cartilage is critical for in vivo efficacy of insulin‐like growth factor 1 in a rat model of osteoarthritis | |
| JP5579445B2 (en) | Peptide compositions and methods for promoting cartilage formation | |
| WO2018186480A1 (en) | Therapeutic medicine for fibrous disease | |
| CN111587117A (en) | Pharmaceutical composition for preventing or treating rheumatoid arthritis containing mitochondria | |
| CN109054030A (en) | A kind of amphoteric ion polymer brush and the preparation method and application thereof based on hyaluronic acid | |
| PT2054050E (en) | Treatment of cartilage disorders with fgf-18 | |
| KR102410986B1 (en) | Fgf-18 compound dosing regimen | |
| CN115088676A (en) | Method for constructing ACPA positive bone erosion rheumatoid arthritis animal model | |
| CN104740603B (en) | Application of polypeptide in preparation of drugs for treatment and/or prevention of rheumatoid arthritis | |
| CN115708872A (en) | Application of anti-angiopoietin-like protein 4 antibody in preparation of medicine for inhibiting expression of inflammatory factor | |
| CN101735314A (en) | Osteopontin and purpose of protecting liver thereof | |
| CN104755095A (en) | Medicament for wound treatment | |
| US20220273695A1 (en) | Mcm for gene therapy to activate wnt pathway | |
| CN114767848A (en) | Method for researching action mechanism of PD-L1 neutralizing antibody on gout inflammation | |
| CN108392624B (en) | Activity promoting peptide and application of mesenchymal stem cells in treating rheumatoid arthritis | |
| JP6489487B2 (en) | Novel use for treating osteoarthritis with tetrapeptide-3 GEKG or pentapeptide-3 GEGF | |
| Taranov et al. | Adjuvant-induced arthritis in guinea pigs | |
| CN111902157B (en) | Composition for promoting local muscle growth, slowing down or preventing local muscle atrophy and application thereof | |
| JP2018528186A (en) | Combination composition comprising FGF-18 compound | |
| Glasson et al. | APPA provides disease modification in preclinical osteoarthritis | |
| Fertala et al. | Prevention of the Post-Traumatic Fibrotic Response in Joints: A Critical Preclinical Evaluation of an Antifibrotic Antibody | |
| Matsuzaki et al. | Range of Motion Exercise Suppresses Myofibroblast Proliferation in the Joint Capsule in a Rat Joint Contracture Model | |
| KR20230030889A (en) | Method of manufacturing an animal model of chronic tophaceous gout |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |