WO2023185448A1 - Use of methyl gallate in preparation of drugs for treating osteoarthritis - Google Patents
Use of methyl gallate in preparation of drugs for treating osteoarthritis Download PDFInfo
- Publication number
- WO2023185448A1 WO2023185448A1 PCT/CN2023/081299 CN2023081299W WO2023185448A1 WO 2023185448 A1 WO2023185448 A1 WO 2023185448A1 CN 2023081299 W CN2023081299 W CN 2023081299W WO 2023185448 A1 WO2023185448 A1 WO 2023185448A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- osteoarthritis
- methyl gallate
- chondrocytes
- preparation
- drugs
- Prior art date
Links
- FBSFWRHWHYMIOG-UHFFFAOYSA-N methyl 3,4,5-trihydroxybenzoate Chemical compound COC(=O)C1=CC(O)=C(O)C(O)=C1 FBSFWRHWHYMIOG-UHFFFAOYSA-N 0.000 title claims abstract description 132
- IBKQQKPQRYUGBJ-UHFFFAOYSA-N methyl gallate Natural products CC(=O)C1=CC(O)=C(O)C(O)=C1 IBKQQKPQRYUGBJ-UHFFFAOYSA-N 0.000 title claims abstract description 65
- 201000008482 osteoarthritis Diseases 0.000 title claims abstract description 54
- 239000003814 drug Substances 0.000 title claims abstract description 26
- 229940079593 drug Drugs 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 210000001612 chondrocyte Anatomy 0.000 claims abstract description 71
- 230000006907 apoptotic process Effects 0.000 claims abstract description 19
- 230000006378 damage Effects 0.000 claims abstract description 10
- 230000007730 Akt signaling Effects 0.000 claims abstract description 8
- 210000000845 cartilage Anatomy 0.000 claims abstract description 8
- 230000000638 stimulation Effects 0.000 claims abstract description 8
- 230000006698 induction Effects 0.000 claims abstract description 5
- 208000027418 Wounds and injury Diseases 0.000 claims abstract 2
- 208000014674 injury Diseases 0.000 claims abstract 2
- 238000011282 treatment Methods 0.000 claims description 13
- 206010061218 Inflammation Diseases 0.000 claims description 10
- 230000004054 inflammatory process Effects 0.000 claims description 10
- 108091008611 Protein Kinase B Proteins 0.000 claims description 8
- 230000037361 pathway Effects 0.000 claims description 7
- 230000004913 activation Effects 0.000 claims description 5
- 210000004027 cell Anatomy 0.000 claims description 5
- 229940126585 therapeutic drug Drugs 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 102000038030 PI3Ks Human genes 0.000 claims 1
- 108091007960 PI3Ks Proteins 0.000 claims 1
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 claims 1
- 102000005765 Proto-Oncogene Proteins c-akt Human genes 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 230000002757 inflammatory effect Effects 0.000 abstract description 8
- 208000037273 Pathologic Processes Diseases 0.000 abstract description 2
- 238000011161 development Methods 0.000 abstract description 2
- 230000005764 inhibitory process Effects 0.000 abstract description 2
- 230000009054 pathological process Effects 0.000 abstract description 2
- 230000035899 viability Effects 0.000 abstract description 2
- 210000002966 serum Anatomy 0.000 description 13
- 241000700159 Rattus Species 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 230000001925 catabolic effect Effects 0.000 description 7
- 238000001262 western blot Methods 0.000 description 7
- 230000003247 decreasing effect Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000000770 proinflammatory effect Effects 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 4
- 101150091111 ACAN gene Proteins 0.000 description 3
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 3
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 3
- 101100096242 Mus musculus Sox9 gene Proteins 0.000 description 3
- 108010087230 Sincalide Proteins 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 2
- 102000051389 ADAMTS5 Human genes 0.000 description 2
- 108091005663 ADAMTS5 Proteins 0.000 description 2
- 101150071146 COX2 gene Proteins 0.000 description 2
- 101100114534 Caenorhabditis elegans ctc-2 gene Proteins 0.000 description 2
- 102100027995 Collagenase 3 Human genes 0.000 description 2
- 101100396994 Drosophila melanogaster Inos gene Proteins 0.000 description 2
- 101000577887 Homo sapiens Collagenase 3 Proteins 0.000 description 2
- 101000990902 Homo sapiens Matrix metalloproteinase-9 Proteins 0.000 description 2
- 102000003777 Interleukin-1 beta Human genes 0.000 description 2
- 108090000193 Interleukin-1 beta Proteins 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 description 2
- 101150000187 PTGS2 gene Proteins 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000003414 extremity Anatomy 0.000 description 2
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000004968 inflammatory condition Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 150000004702 methyl esters Chemical class 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 208000020053 Abnormal inflammatory response Diseases 0.000 description 1
- 206010007710 Cartilage injury Diseases 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 206010018634 Gouty Arthritis Diseases 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- 208000003947 Knee Osteoarthritis Diseases 0.000 description 1
- 102000000874 Pyrin Domain-Containing 3 Protein NLR Family Human genes 0.000 description 1
- 108010001946 Pyrin Domain-Containing 3 Protein NLR Family Proteins 0.000 description 1
- 206010046337 Urate nephropathy Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000003321 cartilage cell Anatomy 0.000 description 1
- 230000008355 cartilage degradation Effects 0.000 description 1
- 230000008367 cartilage synthesis Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000005786 degenerative changes Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229940074391 gallic acid Drugs 0.000 description 1
- 235000004515 gallic acid Nutrition 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000005067 joint tissue Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000378 teratogenic Toxicity 0.000 description 1
- 230000003390 teratogenic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/235—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group
-
- 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
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Definitions
- the present invention relates to the field of medical technology, specifically the application of methyl gallate in the preparation of osteoarthritis therapeutic drugs.
- Osteoarthritis is one of the progressive chronic joint diseases with high disability and teratogenic rates worldwide. It is mainly characterized by the decrease of cartilage cells and the destruction and degradation of joint matrix. Current treatments mainly focus on alleviating clinical symptoms, and there is a lack of effective drugs to improve cartilage degeneration.
- traditional Chinese medicine has the advantages of accurate curative effect and minimal side effects. It has been used clinically, and the recovery of patients after combined treatment with traditional Chinese medicine is significantly better than that of conventional treatment alone. It shows that traditional Chinese medicine has good curative effect in treating osteoarthritis. But how to use traditional Chinese medicine to prevent and treat osteoarthritis is the direction of our research.
- chondrocyte apoptosis may lead to loss of extracellular matrix function and cartilage destruction.
- chondrocyte apoptosis is an inducer of cartilage degradation or a product of cartilage damage
- chondrocyte apoptosis is an important aspect of the pathogenesis of osteoarthritis. Therefore, articular chondrocyte apoptosis and abnormal inflammatory response are one of the initial diseases that lead to degenerative changes in joint tissues. How to alleviate the local inflammatory pressure in the joint and the induced chondrocyte apoptosis has become the key to treating this disease.
- Methyl gallate (Methyl gallate, whose structural formula is shown in Formula I) is a plant phenolic with antioxidant, anti-cancer and anti-inflammatory activities found in a variety of traditional Chinese medicines. It has also been reported that methyl gallate can regulate immune response, inhibit inflammation, and protect blood vessels.
- Chinese patent document CN114028376A discloses the use of MG in the preparation of NLRP3 pathway inhibitors for hyperuricemic nephropathy and/or gouty arthritis.
- Chinese patent document CN108530314A discloses a methyl gallate analog and its application in preparing anti-inflammatory drugs. However, there have been no reports on the application of methyl gallate in preparative osteoarthritis treatment drugs.
- the object of the present invention is to provide the application of methyl gallate in the preparation of osteoarthritis treatment drugs.
- the present invention discovers for the first time that methyl gallate protects chondrocytes by inhibiting the activation of the Pi3k/Akt signaling pathway, inhibiting chondrocyte apoptosis, thereby slowing down and treating osteoarthritis.
- methyl gallate is administered at the same time.
- PCR, Western Blot, and immunofluorescence are used to detect the apoptosis-related Pi3k/Akt signaling pathway, explore the changes in apoptosis expression downstream of Pi3k/Akt, and evaluate the protection of methyl gallate. role of chondrocytes;
- CCK-8 method is used to screen the optimal concentration of methyl gallate intervention
- Methyl gallate inhibits the catabolic ability of chondrocytes in osteoarthritis models
- Methyl gallate alleviates the synthetic ability of chondrocytes in osteoarthritis models
- Methyl gallate down-regulates the Pi3k/Akt pathway and affects chondrocyte function
- the first aspect of the present invention provides the use of methyl gallate in the preparation of osteoarthritis treatment drugs.
- osteoarthritis includes but is not limited to osteoarthritis induced by interleukin-1 ⁇ and/or under conditions stimulated by wind, cold and dampness.
- methyl gallate is used to prepare a medicine for slowing down and treating osteoarthritis caused by inflammation induction and/or damage to chondrocytes caused by wind, cold, and dampness stimulation.
- methyl gallate directly protects chondrocytes by reducing inflammation-induced and/or wind-cold-dampness stimulation damage to chondrocytes, thereby slowing down and treating osteoarthritis.
- methyl gallate protects chondrocytes by inhibiting the activation of the Pi3k/Akt signaling pathway, inhibiting chondrocyte apoptosis, thereby slowing down and treating osteoarthritis.
- a second aspect of the present invention provides the application of methyl gallate in the preparation of a medicine for reducing inflammation-induced and/or wind-cold-dampness stimulation to chondrocyte damage.
- a third aspect of the present invention provides the use of methyl gallate in the preparation of drugs that inhibit the activation of the Pi3k/Akt signaling pathway.
- a fourth aspect of the present invention provides a medicine for treating osteoarthritis, the active ingredient of which is gallic acid. acid methyl ester.
- the medicine also includes pharmaceutically acceptable carriers or excipients.
- the present invention discloses for the first time the protective effect of methyl gallate on chondrocytes in vitro and its application in the treatment of osteoarthritis.
- methyl gallate can reduce the damage of chondrocytes and directly protect chondrocytes; it can also target and inhibit the Pi3k/Akt signaling pathway that controls chondrocyte activation, thereby reducing the release of inflammatory factors.
- methyl gallate can also indirectly protect cartilage by reducing the inflammatory induction of osteoarthritis and/or the inhibition of chondrocyte vitality and apoptosis caused by wind, cold, and dampness stimulation, thereby treating and slowing down osteoarthritis.
- the present invention is supported by a large amount of experimental data, which shows that methyl gallate has a significant protective effect on chondrocytes and can play an important role in the treatment of osteoarthritis in the future.
- the present invention provides new ideas and reference for better understanding the pathological process of osteoarthritis and for the future development of drugs for treating osteoarthritis.
- Figure 1 shows the cytotoxicity test of methyl gallate on rat chondrocytes; A is the 24-hour CCK-8 test to evaluate the cell viability, and B is the 48-hour CCK-8 test to evaluate the cell viability.
- Figure 2 shows the ability of methyl gallate to inhibit the catabolism of chondrocytes in the osteoarthritis model;
- A is the RT-PCR detection of pro-inflammatory factors (IL-6, IL-1 ⁇ ) in SD rat chondrocytes after treatment. ) and catabolic ability (ADAMTS5, MMP13) mRNA expression.
- B is Western Blot to detect the protein expression of iNos, Cox-2 and MMP9 in SD rat chondrocytes after treatment.
- Figure 3 is a diagram showing the synthetic ability of chondrocytes in osteoarthritis model alleviated by methyl gallate;
- A is the immunofluorescence image of Col2 ⁇ 1 and DAPI (nucleus) in each group.
- B shows the fluorescence intensity of Col-2 ⁇ 1 in each group.
- C is Western Blot to detect the protein expression of Col2 ⁇ 1, Acan and Sox-9 in each group.
- Figure 4 is a diagram showing the effects of methyl gallate on the Pi3k/Akt pathway in chondrocytes.
- Figure 5 shows the effect of methyl gallate on chondrocyte apoptosis genes.
- Cartilage from the joints of the limbs of SD rats 24 hours after birth was removed, and chondrocytes were extracted. P1-2 generation cells were studied. CCK8 method was used to detect the cytotoxicity of methyl gallate on rat chondrocytes.
- Example 2 Protective effect of methyl gallate in osteoarthritis
- Cartilage from the joints of the limbs of SD rats 24 hours after birth was removed, and chondrocytes were extracted. P1-2 generation cells were studied.
- Preparation method of wind-cold dampness serum First, use the modified Hulth method + wind-cold-dampness artificial climate chamber intervention to establish a wind-cold-dampness osteoarthritis model (condition setting + humidity 95 ⁇ 2%, temperature 11 ⁇ 1°C, wind level 3 simulated wind-cold dampness environmental factors ), separate rat serum, and inactivate it for later use.
- control group IL-1 ⁇ 20ng/ml
- model group IL-1 ⁇ 20ng/ml
- wind-cold dampness serum group IL-1 ⁇ 20ng/ml+ wind-cold dampness serum 10%
- MG methyl gallate
- Figure 2 is a diagram showing the ability of methyl gallate to inhibit the catabolism of chondrocytes in osteoarthritis.
- pro-inflammatory factors IL-6, IL-1 ⁇
- catabolic indicators ADAMTS5, MMP13, iNos, The expression of Cox-2 and MMP9
- IL-6, IL-1 ⁇ pro-inflammatory factors
- ADAMTS5, MMP13, iNos The expression of Cox-2 and MMP9
- the expression of pro-inflammatory factors and catabolic indicators in the wind-cold-dampness serum group was further increased compared with the model group, indicating that IL-1 ⁇ combined with wind-cold-damp serum intervention can better simulate the inflammatory state of osteoarthritis
- after adding methyl gallate the expression of pro-inflammatory factors and catabolic indicators in chondrocytes significantly decreased, indicating that methyl gallate can inhibit osteoarthritis.
- the catabolic capacity of lower chondrocytes The catabolic capacity of lower chondrocytes.
- Figure 3 is a graph showing the synthetic ability of chondrocytes under the condition of methyl gallate relieving osteoarthritis.
- the results show that the expression of Col2 ⁇ 1, Acan and Sox-9 in the model group decreased compared with the control group, indicating that the chondrocytes after IL-1 ⁇ intervention The cartilage synthesis ability decreased; the expression of Col2 ⁇ 1, Acan and Sox-9 in the wind-cold-dampness serum group decreased significantly compared with the model group, indicating that the chondrocyte synthesis ability was further reduced after the intervention of IL-1 ⁇ combined with wind-cold-dampness serum; and after adding methyl gallate , the expression of synthetic indicators in chondrocytes increased significantly, indicating that methyl gallate can improve the synthetic ability of chondrocytes in osteoarthritis models under inflammatory conditions.
- FIG. 4 is a diagram showing the effects of methyl gallate on the Pi3k/Akt pathway in chondrocytes under osteoarthritis.
- the results show that the expression levels of p-Pi3k and p-AKT in the model group are higher than those in the control group; -The expression levels of Pi3k and p-AKT increased significantly compared with the model group, indicating that the intracellular Pi3k/Akt pathway was activated after the intervention of IL-1 ⁇ combined with wind-cold-dampness serum; and after adding methyl gallate, cartilage
- the expression of p-Pi3k and p-AKT in cells decreased, indicating that methyl gallate regulates the inactivation of the Pi3k/Akt signaling pathway in chondrocytes under inflammatory conditions and affects osteoarthritis.
- Figure 5 shows the effect of methyl gallate on chondrocyte apoptosis genes in osteoarthritis.
- the results show that the expression levels of Bax and Cleave-caspase 3 in the model group are higher than those in the control group, and the expression level of Bcl-2 is higher than that in the control group. group; the expression of Bax and Cleave-caspase3 in the wind-cold-dampness serum group was higher than that of the control group, and the expression of Bcl-2 was lower than that of the control group, indicating that intracellular apoptosis was obvious after the intervention of IL-1 ⁇ combined with wind-cold-dampness serum.
Landscapes
- Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Rheumatology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physical Education & Sports Medicine (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Emergency Medicine (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Pain & Pain Management (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The present invention relates to the technical field of medicines, and in particular, to a use of methyl gallate in the preparation of drugs for treating osteoarthritis. Methyl gallate can relieve the injury of chondrocytes, ameliorate osteoarthritis, and directly protect chondrocytes; the methyl gallate can also inhibit, in a targeted manner, a Pi3k/Akt signaling pathway activated by chondrocytes, thereby reducing the release of inflammatory factors. Besides, the methyl gallate can also play a role in indirectly protecting cartilage by alleviating the inflammatory induction of osteoarthritis and/or chondrocyte viability inhibition and apoptosis caused by wind-cold-damp stimulation, thereby treating and relieving osteoarthritis. In order to better understand the pathological process of osteoarthritis, a new thought and reference are provided for the development of drugs for treating osteoarthritis in the future.
Description
本发明涉及医药技术领域，具体地说，是没食子酸甲酯在制备骨关节炎治疗药物中的应用。The present invention relates to the field of medical technology, specifically the application of methyl gallate in the preparation of osteoarthritis therapeutic drugs.
骨关节炎是目前全球范围内致残率、致畸率较高的进行性慢性关节疾病之一，主要以软骨细胞减少、关节基质破坏降解为特征。目前治疗上主要是以缓解临床症状为主，缺乏有效改善软骨退变的药物。在治疗骨关节炎方面，中医药具有疗效确切、副作用小等优点，已经被应用于临床，并且在联合中药治疗后患者康复情况明显好于单纯性常规治疗。说明中医药在治疗骨关节炎中具有良好的疗效。但如何利用中医药防治骨关节炎是我们研究的方向。Osteoarthritis is one of the progressive chronic joint diseases with high disability and teratogenic rates worldwide. It is mainly characterized by the decrease of cartilage cells and the destruction and degradation of joint matrix. Current treatments mainly focus on alleviating clinical symptoms, and there is a lack of effective drugs to improve cartilage degeneration. In the treatment of osteoarthritis, traditional Chinese medicine has the advantages of accurate curative effect and minimal side effects. It has been used clinically, and the recovery of patients after combined treatment with traditional Chinese medicine is significantly better than that of conventional treatment alone. It shows that traditional Chinese medicine has good curative effect in treating osteoarthritis. But how to use traditional Chinese medicine to prevent and treat osteoarthritis is the direction of our research.
软骨细胞凋亡与膝骨关节炎严重程度之间存在相关性。软骨细胞凋亡可能会导致细胞外基质功能的丧失和软骨破坏。尽管尚不清楚软骨细胞凋亡是软骨退化的诱导剂还是软骨受损后的产物，但软骨细胞凋亡是骨关节炎发病机理的重要方面。因此，关节软骨细胞凋亡及异常炎症反应是导致关节组织发生退行性变化的起始病症之一，如何缓解关节内局部炎症压力及其诱发的软骨细胞凋亡等成为治疗该病症的关键。There is a correlation between chondrocyte apoptosis and severity of knee osteoarthritis. Chondrocyte apoptosis may lead to loss of extracellular matrix function and cartilage destruction. Although it is unclear whether chondrocyte apoptosis is an inducer of cartilage degradation or a product of cartilage damage, chondrocyte apoptosis is an important aspect of the pathogenesis of osteoarthritis. Therefore, articular chondrocyte apoptosis and abnormal inflammatory response are one of the initial diseases that lead to degenerative changes in joint tissues. How to alleviate the local inflammatory pressure in the joint and the induced chondrocyte apoptosis has become the key to treating this disease.
没食子酸甲酯(Methyl gallate，结构式如式I所示)存在与多种中药中，一种具有抗氧化，抗癌和抗炎活性的植物酚类。也有报道没食子酸甲酯可以调节免疫反应、抑制炎症、保护血管。
Methyl gallate (Methyl gallate, whose structural formula is shown in Formula I) is a plant phenolic with antioxidant, anti-cancer and anti-inflammatory activities found in a variety of traditional Chinese medicines. It has also been reported that methyl gallate can regulate immune response, inhibit inflammation, and protect blood vessels.
Methyl gallate (Methyl gallate, whose structural formula is shown in Formula I) is a plant phenolic with antioxidant, anti-cancer and anti-inflammatory activities found in a variety of traditional Chinese medicines. It has also been reported that methyl gallate can regulate immune response, inhibit inflammation, and protect blood vessels.
中国专利文献CN114028376A公开了MG在制备高尿酸血症肾病和/或痛风性关节炎的NLRP3通路抑制剂中的应用。中国专利文献CN108530314A公开了一种没食子酸甲酯类似物及其用于制备抗炎药物的应用。但是关于没食子酸甲酯在制备型骨关节炎治疗药物中的应用目前还未见报道。Chinese patent document CN114028376A discloses the use of MG in the preparation of NLRP3 pathway inhibitors for hyperuricemic nephropathy and/or gouty arthritis. Chinese patent document CN108530314A discloses a methyl gallate analog and its application in preparing anti-inflammatory drugs. However, there have been no reports on the application of methyl gallate in preparative osteoarthritis treatment drugs.
发明内容Contents of the invention
本发明的目的在于提供没食子酸甲酯在制备骨关节炎治疗药物中的应用。本发明首次发现，没食子酸甲酯通过抑制Pi3k/Akt信号通路激活，抑制软骨细胞凋亡而保护软骨细胞，从而减缓和治疗骨关节炎。The object of the present invention is to provide the application of methyl gallate in the preparation of osteoarthritis treatment drugs. The present invention discovers for the first time that methyl gallate protects chondrocytes by inhibiting the activation of the Pi3k/Akt signaling pathway, inhibiting chondrocyte apoptosis, thereby slowing down and treating osteoarthritis.
为了实现上述目的，本发明基于以下技术方案：In order to achieve the above objects, the present invention is based on the following technical solutions:
(1)提取大鼠的原代软骨细胞，检测没食子酸甲酯是否直接减轻炎症下的软骨细胞退变；(1) Extract primary chondrocytes from rats and test whether methyl gallate directly reduces chondrocyte degeneration under inflammation;
(2)炎症诱导后同时给予没食子酸甲酯，用PCR、Western Blot、免疫荧光检测凋亡相关的Pi3k/Akt信号通路，探究Pi3k/Akt下游相关凋亡表达改变情况，评估没食子酸甲酯保护软骨细胞的作用；(2) After inflammation is induced, methyl gallate is administered at the same time. PCR, Western Blot, and immunofluorescence are used to detect the apoptosis-related Pi3k/Akt signaling pathway, explore the changes in apoptosis expression downstream of Pi3k/Akt, and evaluate the protection of methyl gallate. role of chondrocytes;
具体涉及以下实验过程：Specifically involves the following experimental process:
1.原代软骨细胞的提取与鉴定；1. Extraction and identification of primary chondrocytes;
2.CCK-8法筛选没食子酸甲酯干预的最佳浓度；2. CCK-8 method is used to screen the optimal concentration of methyl gallate intervention;
3.没食子酸甲酯抑制骨关节炎模型软骨细胞的分解代谢能力；3. Methyl gallate inhibits the catabolic ability of chondrocytes in osteoarthritis models;
4.没食子酸甲酯缓解骨关节炎模型软骨细胞的合成能力；4. Methyl gallate alleviates the synthetic ability of chondrocytes in osteoarthritis models;
5.没食子酸甲酯下调Pi3k/Akt通路影响软骨细胞功能；5. Methyl gallate down-regulates the Pi3k/Akt pathway and affects chondrocyte function;
6.没食子酸甲酯影响软骨细胞凋亡基因的影响。6. The effect of methyl gallate on chondrocyte apoptosis genes.
基于上述技术方案，本发明的第一方面，提供没食子酸甲酯在制备骨关节炎治疗药物中的应用。Based on the above technical solutions, the first aspect of the present invention provides the use of methyl gallate in the preparation of osteoarthritis treatment drugs.
进一步的，所述的骨关节炎包括但不限于白介素-1β诱导和(或)风寒湿刺激条件下的骨关节炎。Further, the osteoarthritis includes but is not limited to osteoarthritis induced by interleukin-1β and/or under conditions stimulated by wind, cold and dampness.
进一步的，所述的应用中，没食子酸甲酯用于制备减缓和治疗因炎症诱导和(或)风寒湿刺激对软骨细胞的损伤而导致的骨关节炎的药物。Further, in the application, methyl gallate is used to prepare a medicine for slowing down and treating osteoarthritis caused by inflammation induction and/or damage to chondrocytes caused by wind, cold, and dampness stimulation.
进一步的，所述的应用中，没食子酸甲酯通过减轻炎症诱导和(或)风寒湿刺激对软骨细胞的损伤，直接保护软骨细胞，从而减缓和治疗骨关节炎。Further, in the application, methyl gallate directly protects chondrocytes by reducing inflammation-induced and/or wind-cold-dampness stimulation damage to chondrocytes, thereby slowing down and treating osteoarthritis.
进一步的，所述的应用中，没食子酸甲酯通过抑制Pi3k/Akt信号通路激活，抑制软骨细胞凋亡而保护软骨细胞，从而减缓和治疗骨关节炎。Further, in the application described, methyl gallate protects chondrocytes by inhibiting the activation of the Pi3k/Akt signaling pathway, inhibiting chondrocyte apoptosis, thereby slowing down and treating osteoarthritis.
本发明的第二方面，提供没食子酸甲酯在制备减轻炎症诱导和(或)风寒湿刺激对软骨细胞损伤的药物中的应用。A second aspect of the present invention provides the application of methyl gallate in the preparation of a medicine for reducing inflammation-induced and/or wind-cold-dampness stimulation to chondrocyte damage.
本发明的第三方面，提供没食子酸甲酯在制备抑制Pi3k/Akt信号通路激活的药物中的应用。A third aspect of the present invention provides the use of methyl gallate in the preparation of drugs that inhibit the activation of the Pi3k/Akt signaling pathway.
本发明的第四方面，提供一种骨关节炎治疗药物，所述的药物的活性成分为没食子
酸甲酯。A fourth aspect of the present invention provides a medicine for treating osteoarthritis, the active ingredient of which is gallic acid. acid methyl ester.
进一步的，所述的药物还包括药学上可接受的载体或辅料。Furthermore, the medicine also includes pharmaceutically acceptable carriers or excipients.
本发明优点在于：The advantages of the present invention are:
1、本发明首次公开了没食子酸甲酯的保护体外软骨细胞作用及其在治疗骨性关节炎中的应用。1. The present invention discloses for the first time the protective effect of methyl gallate on chondrocytes in vitro and its application in the treatment of osteoarthritis.
2、本发明实验结果证明，没食子酸甲酯可减轻软骨细胞的损伤，直接保护软骨细胞；还可靶向抑制控制软骨细胞激活的Pi3k/Akt信号通路，从而减少炎症因子释放。此外，没食子酸甲酯还可通过减轻骨关节炎的炎性诱导和(或)风寒湿刺激引起的软骨细胞活力抑制和凋亡，起到间接保护软骨的作用，从而治疗和减缓骨关节炎。2. The experimental results of the present invention prove that methyl gallate can reduce the damage of chondrocytes and directly protect chondrocytes; it can also target and inhibit the Pi3k/Akt signaling pathway that controls chondrocyte activation, thereby reducing the release of inflammatory factors. In addition, methyl gallate can also indirectly protect cartilage by reducing the inflammatory induction of osteoarthritis and/or the inhibition of chondrocyte vitality and apoptosis caused by wind, cold, and dampness stimulation, thereby treating and slowing down osteoarthritis.
3、本发明得到了大量实验数据的支持，表明没食子酸甲酯具有显著的软骨细胞保护作用，并可在未来治疗骨关节炎中发挥重要作用。本发明为更好地理解骨关节炎的病理过程，以及今后对治疗骨关节炎药物的开发提供了新的思路与借鉴。3. The present invention is supported by a large amount of experimental data, which shows that methyl gallate has a significant protective effect on chondrocytes and can play an important role in the treatment of osteoarthritis in the future. The present invention provides new ideas and reference for better understanding the pathological process of osteoarthritis and for the future development of drugs for treating osteoarthritis.
图1为没食子酸甲酯对大鼠软骨细胞的细胞毒性检测图；其中，A为24小时CCK-8检测评估细胞的生存能力，B为48小时CCK-8检测评估细胞的生存能力。Figure 1 shows the cytotoxicity test of methyl gallate on rat chondrocytes; A is the 24-hour CCK-8 test to evaluate the cell viability, and B is the 48-hour CCK-8 test to evaluate the cell viability.
图2为没食子酸甲酯抑制骨关节炎模型中软骨细胞的分解代谢能力图；其中，A为RT-PCR检测SD新生大鼠软骨细胞经处理后的促炎因子(IL-6、IL-1β)和分解代谢能力(ADAMTS5、MMP13)的mRNA表达。B为Western Blot检测SD新生大鼠软骨细胞经处理后的iNos、Cox-2和MMP9的蛋白表达。Figure 2 shows the ability of methyl gallate to inhibit the catabolism of chondrocytes in the osteoarthritis model; A is the RT-PCR detection of pro-inflammatory factors (IL-6, IL-1β) in SD rat chondrocytes after treatment. ) and catabolic ability (ADAMTS5, MMP13) mRNA expression. B is Western Blot to detect the protein expression of iNos, Cox-2 and MMP9 in SD rat chondrocytes after treatment.
图3为没食子酸甲酯缓解骨关节炎模型软骨细胞的合成能力图；其中，A为Col2α1与DAPI(细胞核)的各组免疫荧光图像。B为各组的Col-2α1的荧光强度情况。C为Western Blot检测各组的Col2α1、Acan和Sox-9的蛋白表达。Figure 3 is a diagram showing the synthetic ability of chondrocytes in osteoarthritis model alleviated by methyl gallate; A is the immunofluorescence image of Col2α1 and DAPI (nucleus) in each group. B shows the fluorescence intensity of Col-2α1 in each group. C is Western Blot to detect the protein expression of Col2α1, Acan and Sox-9 in each group.
图4为没食子酸甲酯对软骨细胞Pi3k/Akt通路情况图。Figure 4 is a diagram showing the effects of methyl gallate on the Pi3k/Akt pathway in chondrocytes.
图5为没食子酸甲酯对软骨细胞凋亡基因的影响图。Figure 5 shows the effect of methyl gallate on chondrocyte apoptosis genes.
下面结合实施例对本发明提供的具体实施方式作详细说明。The specific implementation modes provided by the present invention will be described in detail below with reference to examples.
实施例1：没食子酸甲酯对大鼠软骨细胞的细胞毒性Example 1: Cytotoxicity of methyl gallate on rat chondrocytes
取出生24h的SD大鼠四肢关节处软骨，提取软骨细胞。P1-2代细胞进行研究。CCK8法检测没食子酸甲酯对大鼠软骨细胞的细胞毒性。Cartilage from the joints of the limbs of SD rats 24 hours after birth was removed, and chondrocytes were extracted. P1-2 generation cells were studied. CCK8 method was used to detect the cytotoxicity of methyl gallate on rat chondrocytes.
不同浓度没食子酸甲酯在不同时间点对软骨细胞活力的影响(图1)。不同浓度没食
子酸甲酯单体干预P1代软骨细胞后，分别于24小时和48小时两个时间点检测450nm处的吸光度值。如图1显示，100μM的没食子酸甲酯干预软骨细胞24、48h时，细胞出现明显生长抑制现象。因此，50μM的没食子酸甲酯对软骨细胞没有毒性。Effects of different concentrations of methyl gallate on chondrocyte viability at different time points (Figure 1). Different concentrations of food After methyl ester monomer interfered with P1 generation chondrocytes, the absorbance value at 450 nm was measured at two time points: 24 hours and 48 hours. As shown in Figure 1, when 100 μM methyl gallate intervened in chondrocytes for 24 and 48 hours, the cells showed obvious growth inhibition. Therefore, 50 μM methyl gallate is not toxic to chondrocytes.
实施例2：没食子酸甲酯在骨关节炎中的保护作用Example 2: Protective effect of methyl gallate in osteoarthritis
取出生24h的SD大鼠四肢关节处软骨，提取软骨细胞。P1-2代细胞进行研究。Cartilage from the joints of the limbs of SD rats 24 hours after birth was removed, and chondrocytes were extracted. P1-2 generation cells were studied.
模拟体外软骨细胞炎症状态：白介素-1β(IL-1β)20ng/ml+风寒湿血清10％共同干预软骨细胞。风寒湿血清制备方法：首先运用改良Hulth法+风寒湿人工气候箱干预建立风寒湿骨关节炎模型(条件设置+为湿度95±2％、温度11±1℃、风力3级模拟风寒湿环境因素)，分离大鼠血清，灭活备用。To simulate the inflammatory state of chondrocytes in vitro: Interleukin-1β (IL-1β) 20ng/ml + wind-cold-dampness serum 10% jointly interfere with chondrocytes. Preparation method of wind-cold dampness serum: First, use the modified Hulth method + wind-cold-dampness artificial climate chamber intervention to establish a wind-cold-dampness osteoarthritis model (condition setting + humidity 95±2%, temperature 11±1℃, wind level 3 simulated wind-cold dampness environmental factors ), separate rat serum, and inactivate it for later use.
设置分组，共计4组：分别是对照组、模型组(IL-1β20ng/ml)、风寒湿血清组(IL-1β20ng/ml+风寒湿血清10％)、没食子酸甲酯(MG)组(IL-1β20ng/ml+风寒湿血清10％+MG50μM)。Set up groups, with a total of 4 groups: control group, model group (IL-1β20ng/ml), wind-cold dampness serum group (IL-1β20ng/ml+ wind-cold dampness serum 10%), and methyl gallate (MG) group (IL- 1β20ng/ml+wind-cold-dampness serum 10%+MG50μM).
收集各组的总RNA和蛋白，进行RT-PCR检测和Western Blot检测软骨细胞的分解代谢指标(图2)。图2为没食子酸甲酯抑制骨关节炎状态下软骨细胞的分解代谢能力图，结果显示：模型组的促炎因子(IL-6、IL-1β)、分解代谢指标(ADAMTS5、MMP13、iNos、Cox-2和MMP9)的表达量较对照组升高，说明IL-1β干预后的软骨细胞出现炎症状态；风寒湿血清组的促炎因子和分解代谢指标的表达较模型组进一步升高，说明IL-1β联合风寒湿血清干预更能模拟骨关节炎的炎症状态；而加入没食子酸甲酯后，软骨细胞中促炎因子和分解代谢指标表达明显下降，说明没食子酸甲酯能够抑制骨关节炎下软骨细胞的分解代谢能力。Total RNA and protein from each group were collected, and RT-PCR detection and Western Blot were performed to detect the catabolic indicators of chondrocytes (Figure 2). Figure 2 is a diagram showing the ability of methyl gallate to inhibit the catabolism of chondrocytes in osteoarthritis. The results show: pro-inflammatory factors (IL-6, IL-1β), catabolic indicators (ADAMTS5, MMP13, iNos, The expression of Cox-2 and MMP9) was increased compared with the control group, indicating that chondrocytes after IL-1β intervention were in an inflammatory state; the expression of pro-inflammatory factors and catabolic indicators in the wind-cold-dampness serum group was further increased compared with the model group, indicating that IL-1β combined with wind-cold-damp serum intervention can better simulate the inflammatory state of osteoarthritis; and after adding methyl gallate, the expression of pro-inflammatory factors and catabolic indicators in chondrocytes significantly decreased, indicating that methyl gallate can inhibit osteoarthritis. The catabolic capacity of lower chondrocytes.
免疫荧光和Western Blot检测软骨细胞的合成能力指标(图3)。图3为没食子酸甲酯缓解骨关节炎状态下软骨细胞的合成能力图，结果显示，模型组的Col2α1、Acan和Sox-9的表达量较对照组下降，说明IL-1β干预后的软骨细胞出现软骨合成能力下降；风寒湿血清组的Col2α1、Acan和Sox-9表达量较模型组下降明显，说明IL-1β联合风寒湿血清干预后软骨细胞合成能力进一步降低；而加入没食子酸甲酯后，软骨细胞中合成指标表达明显上升，说明没食子酸甲酯能够改善炎症状态下骨关节炎模型中软骨细胞的合成能力。Immunofluorescence and Western Blot were used to detect the synthetic ability indicators of chondrocytes (Figure 3). Figure 3 is a graph showing the synthetic ability of chondrocytes under the condition of methyl gallate relieving osteoarthritis. The results show that the expression of Col2α1, Acan and Sox-9 in the model group decreased compared with the control group, indicating that the chondrocytes after IL-1β intervention The cartilage synthesis ability decreased; the expression of Col2α1, Acan and Sox-9 in the wind-cold-dampness serum group decreased significantly compared with the model group, indicating that the chondrocyte synthesis ability was further reduced after the intervention of IL-1β combined with wind-cold-dampness serum; and after adding methyl gallate , the expression of synthetic indicators in chondrocytes increased significantly, indicating that methyl gallate can improve the synthetic ability of chondrocytes in osteoarthritis models under inflammatory conditions.
Western Blot检测Pi3k/Akt通路蛋白水平(图4)。图4为没食子酸甲酯对骨关节炎状态下软骨细胞Pi3k/Akt通路情况图，结果显示，模型组的p-Pi3k、p-AKT的表达量较对照组升高；风寒湿血清组的p-Pi3k、p-AKT的表达量较模型组上升明显，说明IL-1β联合风寒湿血清干预后的细胞内Pi3k/Akt通路被激活；而加入没食子酸甲酯后，软骨
细胞中p-Pi3k、p-AKT的表达量下降，说明没食子酸甲酯调控炎症状态下的软骨细胞中Pi3k/Akt信号通路失活影响骨关节炎。Western Blot detected Pi3k/Akt pathway protein levels (Figure 4). Figure 4 is a diagram showing the effects of methyl gallate on the Pi3k/Akt pathway in chondrocytes under osteoarthritis. The results show that the expression levels of p-Pi3k and p-AKT in the model group are higher than those in the control group; -The expression levels of Pi3k and p-AKT increased significantly compared with the model group, indicating that the intracellular Pi3k/Akt pathway was activated after the intervention of IL-1β combined with wind-cold-dampness serum; and after adding methyl gallate, cartilage The expression of p-Pi3k and p-AKT in cells decreased, indicating that methyl gallate regulates the inactivation of the Pi3k/Akt signaling pathway in chondrocytes under inflammatory conditions and affects osteoarthritis.
Western Blot检测软骨细胞凋亡蛋白水平(图5)。图5为没食子酸甲酯对骨关节炎状态下软骨细胞凋亡基因的影响图，结果显示，模型组的Bax、Cleave-caspase 3的表达量较对照组升高，Bcl-2表达量较对照组降低；风寒湿血清组的Bax、Cleave-caspase3的表达量较对照组升高，Bcl-2表达量较对照组更低，说明IL-1β联合风寒湿血清干预后的细胞内凋亡情况明显加剧；而加入没食子酸甲酯后，软骨细胞中Bax、Cleave-caspase3的表达量较对照组降低，Bcl-2表达量较对照组升高，说明没食子酸甲酯降低炎症状态下软骨细胞中凋亡基因的表达，从而在骨关节炎起保护作用。Western Blot detected chondrocyte apoptosis protein levels (Figure 5). Figure 5 shows the effect of methyl gallate on chondrocyte apoptosis genes in osteoarthritis. The results show that the expression levels of Bax and Cleave-caspase 3 in the model group are higher than those in the control group, and the expression level of Bcl-2 is higher than that in the control group. group; the expression of Bax and Cleave-caspase3 in the wind-cold-dampness serum group was higher than that of the control group, and the expression of Bcl-2 was lower than that of the control group, indicating that intracellular apoptosis was obvious after the intervention of IL-1β combined with wind-cold-dampness serum. After adding methyl gallate, the expression of Bax and Cleave-caspase3 in chondrocytes was lower than that of the control group, and the expression of Bcl-2 was higher than that of the control group, indicating that methyl gallate reduces apoptosis in chondrocytes under inflammation. expression of death genes, thus playing a protective role in osteoarthritis.
以上已对本发明创造的较佳实施例进行了具体说明，但本发明创造并不限于所述实施例，熟悉本领域的技术人员在不违背本发明创造精神的前提下还可做出种种的等同的变型或替换，这些等同的变型或替换均包含在本申请权利要求所限定的范围内。
The preferred embodiments of the invention have been specifically described above, but the invention is not limited to the embodiments. Those skilled in the art can also make various equivalents without violating the spirit of the invention. modifications or substitutions, these equivalent modifications or substitutions are included in the scope defined by the claims of this application.
Claims (8)
- 没食子酸甲酯在制备骨关节炎治疗药物中的应用。Application of methyl gallate in the preparation of osteoarthritis treatment drugs.
- 根据权利要求1所述的没食子酸甲酯在制备骨关节炎治疗药物中的应用，其特征在于，没食子酸甲酯用于制备减缓和治疗因炎症诱导和(或)风寒湿刺激对软骨细胞的损伤而导致的骨关节炎的药物。The application of methyl gallate in the preparation of osteoarthritis therapeutic drugs according to claim 1, characterized in that, methyl gallate is used to prepare and slow down and treat the damage to chondrocytes caused by inflammation induction and/or wind, cold and dampness stimulation. Drugs for osteoarthritis caused by injury.
- 根据权利要求1所述的没食子酸甲酯在制备骨关节炎治疗药物中的应用，其特征在于，没食子酸甲酯通过减轻炎症诱导和(或)风寒湿刺激对软骨细胞的损伤，直接保护软骨细胞，从而减缓和治疗骨关节炎。The application of methyl gallate in the preparation of osteoarthritis therapeutic drugs according to claim 1, characterized in that methyl gallate directly protects cartilage by reducing inflammation induction and/or wind-cold-dampness stimulation to chondrocytes. cells, thereby slowing and treating osteoarthritis.
- 根据权利要求1所述的没食子酸甲酯在制备骨关节炎治疗药物中的应用，其特征在于，没食子酸甲酯通过抑制磷脂酰肌醇-3-激酶/蛋白激酶B(Pi3k/Akt)信号通路激活，抑制软骨细胞凋亡而保护软骨细胞，从而减缓和治疗骨关节炎。The application of methyl gallate in the preparation of osteoarthritis therapeutic drugs according to claim 1, characterized in that, methyl gallate inhibits phosphatidylinositol-3-kinase/protein kinase B (Pi3k/Akt) signal The pathway is activated to inhibit chondrocyte apoptosis and protect chondrocytes, thereby slowing down and treating osteoarthritis.
- 没食子酸甲酯在制备减轻炎症诱导和(或)风寒湿刺激对软骨细胞损伤的药物中的应用。Application of methyl gallate in the preparation of drugs for reducing chondrocyte damage induced by inflammation and/or wind, cold, and dampness stimulation.
- 没食子酸甲酯在制备抑制Pi3k/Akt信号通路激活的药物中的应用。Application of methyl gallate in the preparation of drugs that inhibit the activation of Pi3k/Akt signaling pathway.
- 一种骨关节炎治疗药物，其特征在于，所述的药物的活性成分为没食子酸甲酯。A medicine for treating osteoarthritis, characterized in that the active ingredient of the medicine is methyl gallate.
- 根据权利要求7所述的骨关节炎治疗药物，其特征在于，所述的药物还包括药学上可接受的载体或辅料。 The osteoarthritis treatment drug according to claim 7, characterized in that the drug further includes a pharmaceutically acceptable carrier or auxiliary material.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210322511.X | 2022-03-30 | ||
| CN202210322511.XA CN114931571A (en) | 2022-03-30 | 2022-03-30 | Application of gallic acid methyl ester in preparation of osteoarthritis treatment medicine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2023185448A1 true WO2023185448A1 (en) | 2023-10-05 |
Family
ID=82861570
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2023/081299 WO2023185448A1 (en) | 2022-03-30 | 2023-03-14 | Use of methyl gallate in preparation of drugs for treating osteoarthritis |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN114931571A (en) |
| WO (1) | WO2023185448A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114931571A (en) * | 2022-03-30 | 2022-08-23 | 上海中医药大学附属曙光医院 | Application of gallic acid methyl ester in preparation of osteoarthritis treatment medicine |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104884067A (en) * | 2012-10-17 | 2015-09-02 | 甲基化物科学国际有限公司 | Compositions comprising S-adenosylmethionine and a gallic acid ester |
| CN108530314A (en) * | 2018-05-23 | 2018-09-14 | 温州医科大学 | A kind of gallicin analog and its application |
| WO2020050602A1 (en) * | 2018-09-05 | 2020-03-12 | (주)루젠에스씨아이 | Screening system for human-transformed cartilage cell line-based cartilage disease treatment agents |
| CN114931571A (en) * | 2022-03-30 | 2022-08-23 | 上海中医药大学附属曙光医院 | Application of gallic acid methyl ester in preparation of osteoarthritis treatment medicine |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2013134126A (en) * | 2011-01-10 | 2015-02-20 | ЕэСэМ ТЕКНОЛОДЖИС, ЛЛК | METHOD FOR REDUCING THE LEVEL OF THE C-END BODY COLLAGEN TYPE 2 |
| KR101401154B1 (en) * | 2011-02-22 | 2014-05-30 | 경희대학교 산학협력단 | Pharmaceutical composition for inhibiting regulatory T cell activity comprising methyl gallate |
| CN108863892B (en) * | 2018-07-04 | 2022-04-05 | 温州医科大学 | Gallic acid methyl ester analogue containing amide structure and application thereof |
| CN114028376A (en) * | 2021-09-17 | 2022-02-11 | 武汉翼博济生生物科技有限公司 | Application of MG in preparation of NLRP3 pathway inhibitor for hyperuricemia nephropathy and/or gouty arthritis |
-
2022
- 2022-03-30 CN CN202210322511.XA patent/CN114931571A/en active Pending
-
2023
- 2023-03-14 WO PCT/CN2023/081299 patent/WO2023185448A1/en unknown
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104884067A (en) * | 2012-10-17 | 2015-09-02 | 甲基化物科学国际有限公司 | Compositions comprising S-adenosylmethionine and a gallic acid ester |
| CN108530314A (en) * | 2018-05-23 | 2018-09-14 | 温州医科大学 | A kind of gallicin analog and its application |
| WO2020050602A1 (en) * | 2018-09-05 | 2020-03-12 | (주)루젠에스씨아이 | Screening system for human-transformed cartilage cell line-based cartilage disease treatment agents |
| CN114931571A (en) * | 2022-03-30 | 2022-08-23 | 上海中医药大学附属曙光医院 | Application of gallic acid methyl ester in preparation of osteoarthritis treatment medicine |
Non-Patent Citations (5)
| Title |
|---|
| CORREA LUANA BARBOSA, PÁDUA TATIANA ALMEIDA, SEITO LEONARDO NOBORU, COSTA THADEU ESTEVAM MOREIRA MARAMALDO, SILVA MAGAIVER ANDRADE: "Anti-inflammatory Effect of Methyl Gallate on Experimental Arthritis: Inhibition of Neutrophil Recruitment, Production of Inflammatory Mediators, and Activation of Macrophages", JOURNAL OF NATURAL PRODUCTS, AMERICAN CHEMICAL SOCIETY, US, vol. 79, no. 6, 24 June 2016 (2016-06-24), US , pages 1554 - 1566, XP093094445, ISSN: 0163-3864, DOI: 10.1021/acs.jnatprod.5b01115 * |
| CORREA, LUANA BARBOSA ET AL.: "Protective effect of methyl gallate on murine antigen‑induced arthritis by inhibiting inflammatory process and bone erosion", INFLAMMOPHARMACOLOGY, vol. 30, 2 February 2022 (2022-02-02), pages 251 - 266, XP037714779, DOI: 10.1007/s10787-021-00922-8 * |
| WENZHOU SCIENCE & TECHNOLOGY BUREAU: "Notice of Wenzhou Science and Technology Bureau on the Establishment of Wenzhou Basic Scientific Research Project (self-funded) in 2021", WENZHOU SCIENCE & TECHNOLOGY BUREAU > GOVERNMENT AFFAIRS DISCLOSURE > TECHNOLOGY PROJECTS - NOTICES, WENZHOU SCIENCE & TECHNOLOGY BUREAU, CN, CN, XP009549989, Retrieved from the Internet <URL:https://wzkj.wenzhou.gov.cn/art/2021/8/13/art_1220139_58895056.html> * |
| XIAN-BAO ZHANG, WANG YUN: "Mechanism Elucidation of Euphorbia helioscopia for Treatment of Lung Cancer Based on Entity Grammar System", MODERN CHINESE MEDICINE, ZHONG GUO XIAN DAI ZHONG YAO BIAN JI BU, CN, vol. 23, no. 2, 26 March 2021 (2021-03-26), CN , pages 294 - 302, XP093094777, ISSN: 1673-4890, DOI: 10.13313/j.issn.1673-4890.20200324003 * |
| XUE SHEN, XUE YAN, CHEN YAN, LI MING, ZHANG HU: "Establishment and Evaluation of Osteoarthritis Model in Vivo and in Vitro Under the Guidance of The Theory of "Wind-Cold-Dampness Causing Arthralgia", JOURNAL OF BASIC CHINESE MEDICINE, vol. 27, no. 11, 4 February 2021 (2021-02-04), pages 1721 - 1724, XP093094778, DOI: 10.19945/j.cnki.issn.1006-3250.2021.11.010 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114931571A (en) | 2022-08-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Rosillo et al. | Protective effect of ellagic acid, a natural polyphenolic compound, in a murine model of Crohn's disease | |
| Wu et al. | Geraniol-mediated osteoarthritis improvement by down-regulating PI3K/Akt/NF-κB and MAPK signals: In vivo and in vitro studies | |
| WO2023185448A1 (en) | Use of methyl gallate in preparation of drugs for treating osteoarthritis | |
| CN109453173A (en) | The purposes of iron death inhibitor | |
| WO2005082349A1 (en) | Composition for the treatment of obesity comprising fumagillol derivative | |
| Shen et al. | Morphological and functional changes of mitochondria in apoptotic esophageal carcinoma cells induced by arsenic trioxide | |
| Chen et al. | Regulation and therapy, the role of JAK2/STAT3 signaling pathway in OA: a systematic review | |
| CN112823796A (en) | Application of protein tyrosine phosphatase SHP2 inhibitor in preparation of medicine for treating osteoarthritis | |
| CN112870361A (en) | Use of iron death inhibitor for preparing medicine for preventing or treating osteoporosis or bone loss caused by iron overload | |
| Wang et al. | Potential therapeutic role of Co-Q10 in alleviating intervertebral disc degeneration and suppressing IL-1β-mediated inflammatory reaction in NP cells | |
| Fu et al. | Grape seed proanthocyanidins attenuate apoptosis in ischemic stroke | |
| Wang et al. | Oxymatrine liposomes for intervertebral disc treatment: formulation, in vitro and vivo assessments | |
| Yan et al. | Rhoifolin ameliorates osteoarthritis via regulating autophagy | |
| Liu et al. | Chebulanin exerts its anti-inflammatory and anti-arthritic effects via inhibiting NF-κB and MAPK activation in collagen-induced arthritis mice | |
| Zhang et al. | 18β-Glycyrrhetinic acid monoglucuronide (GAMG) alleviates single-walled carbon nanotubes (SWCNT)-induced lung inflammation and fibrosis in mice through PI3K/AKT/NF-κB signaling pathway | |
| Wang et al. | Mangiferin exert protective effects on joints of adjuvant-induced arthritis rats by regulating the MAPKs/NF-κB pathway of fibroblast-like synoviocytes | |
| Shen et al. | Si Miao San attenuates inflammation and oxidative stress in rats with CIA via the modulation of the Nrf2/ARE/PTEN pathway | |
| Cheng et al. | Protective effects of hydrogen-rich water against cartilage damage in a rat model of osteoarthritis by inhibiting oxidative stress, matrix catabolism, and apoptosis | |
| Zhang et al. | Edaravone Dexborneol Alleviates Cerebral Ischemic Injury via MKP‐1‐Mediated Inhibition of MAPKs and Activation of Nrf2 | |
| Hong et al. | Total flavonoids of bidens pilosa ameliorates bone destruction in collagen-induced arthritis | |
| Huang et al. | The disease-modifying effect of dehydroepiandrosterone in different stages of experimentally induced osteoarthritis: a histomorphometric study | |
| Shi et al. | κ-Opioid receptor activation attenuates osteoarthritis synovitis by regulating macrophage polarization through the NF-κB pathway: KOR regulates OA synovitis via NF-κB | |
| Xu et al. | Benzalkonium bromide as a new potential instillation drug for bladder cancer: hypothesis and pilot study | |
| Kim et al. | Early and synergistic recovery effect of herbal combination on surgically corrected varicocele | |
| Jin et al. | Corilagin attenuates airway inflammation and collagen deposition in ovalbumin-induced asthmatic mice |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 23777824 Country of ref document: EP Kind code of ref document: A1 |