Application of adenosine monophosphate in preparation of medicine for treating osteoarthritis
By using adenosine monophosphate solution to inhibit the expression of IL-6 and MMP13 in osteoarthritis chondrocytes, cartilage damage was reduced, solving the problem that existing drugs could not improve cartilage degeneration, and achieving early intervention and disease improvement in osteoarthritis.
Patent Information
- Application Number
- CN202511932108.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-19
- Publication Date
- 2026-01-23
AI Technical Summary
Current medications for osteoarthritis are ineffective in improving articular cartilage degeneration, primarily relieving pain but failing to reverse disease progression.
The protein expression of inflammatory factors IL-6 and matrix metalloproteinase MMP13 in chondrocytes was inhibited by 1 mM to 2 mM adenosine monophosphate solution, and cartilage damage was alleviated by intra-articular injection of 1 mg/mL adenosine monophosphate saline solution.
It significantly reduces the degree of cartilage damage, providing a new way to improve the condition of osteoarthritis by inhibiting joint inflammation and chondrocyte degradation mediated by key factors, thus achieving early intervention.
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Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biological medicine, and particularly relates to application of adenosine monophosphate in preparation of a drug for treating osteoarthritis. BACKGROUND
[0002] Osteoarthritis is a common chronic inflammatory bone joint disabling disease, which often leads to significant decline in limb function, life quality and exercise level of patients. The main pathological changes of osteoarthritis include joint cartilage degeneration, synovial inflammation and abnormal bone remodeling. At present, the pathological process and mechanism of osteoarthritis have not been fully elucidated, and the recognized view is that the occurrence and development of osteoarthritis are closely related to the joint cartilage degeneration, cartilage cell apoptosis and extracellular matrix degradation caused by inflammation and immune factors. As a physiological structure contacting with bones, cartilage plays a role in weight bearing, stress transduction and bone structure buffering. Because of its important physiological position and function, the damage and degeneration of cartilage is the main reason for the weakening or loss of joint function, and is also the core pathological feature of the occurrence and development of osteoarthritis.
[0003] At present, the osteoarthritis treatment drugs mainly relieve clinical symptoms and alleviate pain of patients, and are difficult to improve cartilage degeneration. Therefore, it is necessary to explore a new way for improving osteoarthritis. SUMMARY
[0004] In order to explore a new way for improving osteoarthritis, the application provides application of adenosine monophosphate in preparation of a drug for treating osteoarthritis. The 1 mM-2 mM adenosine monophosphate solution can dose-dependently inhibit protein expression of inflammatory factors IL-6 and matrix metalloproteinase MMP13 in cartilage cells, and injection of 1 mg / mL adenosine monophosphate physiological saline solution can significantly reduce the degree of cartilage damage, thereby providing a new way for improving osteoarthritis.
[0005] The application provides application of adenosine monophosphate in preparation of a drug for treating osteoarthritis, wherein the adenosine monophosphate has a molecular formula of C 10 H 14 The chemical structural formula of the adenosine monophosphate is as follows: .
[0006] The 1 mM-2 mM adenosine monophosphate solution can dose-dependently inhibit protein expression of inflammatory factors IL-6 and matrix metalloproteinase MMP13 in cartilage cells, and injection of 1 mg / mL adenosine monophosphate physiological saline solution can significantly reduce the degree of cartilage damage, thereby providing a new way for improving osteoarthritis.
[0007] Further, the drug is an injection.
[0008] Further, the medicine is prepared from the adenosine monophosphate and an excipient.
[0009] Further, the excipient is physiological saline.
[0010] Further, the injection is an adenosine monophosphate physiological saline solution with a concentration of 0.2 mg / mL to 1 mg / mL.
[0011] Further, the medicine is used for relieving cartilage damage of osteoarthritis.
[0012] Further, the medicine is used for inhibiting expression of inflammatory factor IL-6 protein.
[0013] Further, the medicine is used for inhibiting expression of matrix metalloproteinase MMP13 protein.
[0014] Compared with the prior art, the present application has the following beneficial effects: The adenosine monophosphate solution in the present application can simultaneously inhibit protein expression of inflammatory factor IL-6 and matrix metalloproteinase MMP13 in chondrocytes, and early intervention is performed on cartilage degeneration caused by osteoarthritis by inhibiting key factors mediating arthritis and chondrocyte degradation; the experiment of an iodine acetic acid-induced rat osteoarthritis model shows that after intra-articular injection of 1 mg / mL adenosine monophosphate physiological saline solution, the degree of cartilage damage is significantly reduced compared with the model group, proving that adenosine monophosphate has the function of reversing cartilage degeneration, and providing a new way for improvement of osteoarthritis. BRIEF DESCRIPTION OF DRAWINGS
[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description only some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of these drawings.
[0016] Figure 1 is CCK-8 detection of the influence of adenosine monophosphate on the activity of SW1353 cells.
[0017] Figure 2 is ELISA detection of the influence of adenosine monophosphate on the expression of IL-6 protein in SW1353 cells; in the figure, - means no addition, and + means addition.
[0018] Figure 3 is ELISA detection of the influence of adenosine monophosphate on the expression of MMP13 protein in SW1353 cells; in the figure, - means no addition, and + means addition.
[0019] Figure 4Effect of adenosine monophosphate on cartilage degeneration of rat knee osteoarthritis In the figure, A is the introduction of the test treatment time; B is the photo of the damage degree of the knee osteoarthritis cartilage surface of the rats in different treatment groups. DETAILED DESCRIPTION
[0020] The specific embodiments of the present application are described in detail below, but it should be understood that the scope of protection of the present application is not limited by the specific embodiments. Based on the examples in the present application, all other examples obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application. The experimental methods described in the embodiments of the present application are conventional methods, and the materials and reagents used in the following examples are commercially available unless otherwise specified.
[0021] Example 1: Application of adenosine monophosphate in the preparation of a drug for treating osteoarthritis.
[0022] I. Test materials Adenosine monophosphate was purchased from Guangzhou He Wei Pharmaceutical Technology Co., Ltd. CD105311-25 g.
[0023] SW1353 cells were purchased from Wuhan Punsai Life Science Co., Ltd. SW1353 cells have good cartilage phenotype and are suitable for research on cartilage cell structure and function.
[0024] II. Test method 1. Effect of adenosine monophosphate on the viability of SW1353 cells Prepare a SW1353 cell suspension with a concentration of 5×10 4 SW1353 cells were prepared at a concentration of 5×10
[0025] Dissolve adenosine monophosphate in DMEM to prepare a high-concentration stock solution (100 mM), then take 0.5 μL, 1 μL, 2 μL, 4 μL, and 10 μL of the high-concentration stock solution and add them to 100 μL of complete culture medium (DMEM culture medium containing 10% by volume of serum) to obtain 0.5 mM, 1 mM, 2 mM, 4 mM, and 10 mM adenosine monophosphate solutions.
[0026] Control and treatment groups were set up. The control group was given 100 μL of fresh complete culture medium, while the treatment groups were given 100 μL of 0.5 mM, 1 mM, 2 mM, 4 mM, and 10 mM adenosine monophosphate solutions, respectively. After treatment, both control and treatment groups were cultured at 37℃ and 5% CO2 for 24 h, and the viability of SW1353 cells was detected using a CCK8 assay (Cell Counting Kit-8, purchased from Shanghai Huaxiang Biotechnology Co., Ltd.).
[0027] 2. Effect of adenosine monophosphate on IL-6 protein expression in SW1353 cells Prepare a cell concentration of 6 × 10⁶ 4 SW1353 cell suspension was prepared by layering the SW1353 cell suspension into 24-well plates, with 500 μL added to each well. After 36 h, the medium was replaced with DMEM medium containing 1% serum for serum starvation.
[0028] SW1353 cells starved for 24 h were divided into control group 1, control group 2, and treatment group. The treatment group was pretreated with 1 mM and 2 mM adenosine monophosphate (ATP) solutions for 3 h, followed by the addition of the inflammatory stimulant IL-1β to achieve a final IL-1β concentration of 10 ng / mL. Control group 2 was not treated with ATP solution, but was pretreated under the same conditions for 3 h before being supplemented with IL-1β to achieve a final IL-1β concentration of 10 ng / mL. Control group 1 received neither IL-1β nor ATP solution. After treatment at 37℃ and 5% CO2 for 24 h, the protein expression level of the inflammatory cytokine IL-6 was detected by ELISA (Human IL-6 ELISA Kit, purchased from Wuhan Sanying Biotechnology Co., Ltd.).
[0029] 3. Effect of adenosine monophosphate on MMP13 protein expression in SW1353 cells Prepare a cell concentration of 6 × 10⁶ 4 SW1353 cell suspension was prepared by layering the SW1353 cell suspension into 24-well plates, with 500 μL added to each well. After 36 h, the medium was replaced with DMEM medium containing 1% serum for serum starvation.
[0030] SW1353 cells after serum starvation for 24 h were divided into control group 1, control group 2 and treatment group. The treatment group was pre-treated with 1 mM and 2 mM adenosine monophosphate solution for 3 h, and then inflammatory stimulant IL-1β was added to make the final concentration of IL-1β 10 ng / mL; the control group 2 was not added with adenosine monophosphate solution, and was pre-treated under the same conditions for 3 h, and then inflammatory stimulant IL-1β was added to make the final concentration of IL-1β 10 ng / mL; the control group 1 was not added with IL-1β and adenosine monophosphate solution. After the three groups of cells were treated at 37℃, 5% CO2 for 24 h, the protein expression level of inflammatory factor MMP13 of the cells was detected by ELISA (Human MMP13 ELISA Kit, purchased from Wuhan Sany Biological Technology Co., Ltd.).
[0031] 4. Effect of adenosine monophosphate on cartilage degeneration of rat knee osteoarthritis induced by iodoacetate After adaptive feeding for 7 days, 200±20 g male 6-week-old SD rats were randomly divided into a control group, a model group, a drug administration group one and a drug administration group two, a total of 4 groups.
[0032] 10 mg of adenosine monophosphate was weighed and dissolved in 50 mL of a physiological saline solution to prepare a 0.2 mg / mL adenosine monophosphate physiological saline solution; 50 mg of adenosine monophosphate was weighed and dissolved in 50 mL of a physiological saline solution to prepare a 1 mg / mL adenosine monophosphate physiological saline solution.
[0033] On day 1, the drug administration group one was injected with the 0.2 mg / mL adenosine monophosphate physiological saline solution in the right knee joint cavity of the rats, 50 μL per rat; the drug administration group two was injected with the 1 mg / mL adenosine monophosphate physiological saline solution in the right knee joint cavity of the rats, 50 μL per rat; the model group and the control group were injected with the same amount of physiological saline in the right knee joint cavity of the rats.
[0034] On day 2, the model group, the drug administration group one and the drug administration group two were injected with the 20 mg / mL iodoacetate solution in the right knee joint cavity of the rats, 50 μL per rat, to construct the knee osteoarthritis model; the control group was injected with the same amount of physiological saline in the right knee joint cavity of the rats.
[0035] On days 3, 5, 7, 9, 11 and 13, the drug administration group one was injected with the 0.2 mg / mL adenosine monophosphate physiological saline solution in the right knee joint cavity of the rats, 50 μL per rat; the drug administration group two was injected with the 1 mg / mL adenosine monophosphate physiological saline solution in the right knee joint cavity of the rats, 50 μL per rat; the control group and the model group were injected with the same amount of physiological saline in the right knee joint cavity of the rats.
[0036] On the 15th day, the right knee joint samples of the control group, the model group, the first drug administration group and the second drug administration group were collected, and the right knee joint cartilage damage of the rats in each group was observed.
[0037] III. Test Results 1. Adenosine monophosphate inhibits SW1353 cell viability The CCK8 test results of SW1353 cells are shown in Table 1. Figure 1 When the final concentration of adenosine monophosphate added was 0.5 mM, 1 mM, 2 mM, 4 mM and 10 mM, the OD values were all significantly different from those of the control group (P<0.05), but there was no difference between these concentrations (P>0.05), indicating that adenosine monophosphate had a certain inhibitory effect on the viability of SW1353 cells. 450
[0038] 2. Adenosine monophosphate inhibits the expression of IL-6 and MMP13 proteins in SW1353 cells IL-1β was used as an inflammatory stimulant to construct an osteoarthritis model of SW1353 cells in the present application. The ELISA test results showed that, compared with the control group 2 (only IL-1β was added), the protein content of IL-6 in the treatment group (adenosine monophosphate solution and IL-1β were added) gradually decreased with the increase of the concentration of adenosine monophosphate solution, and there was a significant difference (P<0.05) between the treatment group and the control group 2 (Table 2). Figure 2 Compared with the control group 2 (only IL-1β was added), the protein content of MMP13 in the treatment group (adenosine monophosphate solution and IL-1β were added) gradually decreased with the increase of the concentration of adenosine monophosphate solution, and there was a significant difference (P<0.05) between the treatment group and the control group 2 (Table 3). Figure 3 This indicates that adenosine monophosphate can inhibit the expression of inflammatory factors IL-6 and matrix metalloproteinase MMP13 proteins.
[0039] 3. Adenosine monophosphate alleviates iodine acetic acid-induced cartilage degeneration of rat knee osteoarthritis The test processing scheme is shown in Figure A. Figure 4 The test results showed that, compared with the control group, the knee joint cartilage of the model group of rats injected with 20 mg / mL iodine acetic acid solution was obviously damaged, indicating that the rat knee osteoarthritis model was successfully constructed; and compared with the model group, the degree of knee joint cartilage damage of the rats in the second drug administration group added with 1 mg / mL normal saline solution was significantly reduced (Figure B), indicating that adenosine monophosphate can alleviate iodine acetic acid-induced cartilage degeneration of rat knee osteoarthritis. Figure 4
[0040] Although the preferred embodiments of the present application have been described, those skilled in the art can make further changes and modifications to these embodiments once they know the basic inventive concept.
[0041] It will be apparent to those skilled in the art that various modifications and variations can be made to the present application without departing from the spirit or scope of the application. Thus, it is intended that the present application cover modifications and variations of this application provided they come within the scope of the appended claims and their equivalents.
Claims
1. Use of adenosine monophosphate for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The adenosine monophosphate has a molecular formula of C 10 H 14 N5O7P, and a chemical structural formula as shown in the following. 。 2. Use of adenosine monophosphate according to claim 1 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The medicine is an injection.
3. Use of adenosine monophosphate according to claim 2 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The medicine is prepared from the adenosine monophosphate and an excipient.
4. Use of adenosine monophosphate according to claim 2 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The excipient is physiological saline.
5. Use of adenosine monophosphate according to claim 2 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The injection is an adenosine monophosphate physiological saline solution with a concentration of 0.2 mg / mL to 1 mg / mL.
6. Use of adenosine monophosphate according to claim 1 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The medicine is used for relieving cartilage damage of osteoarthritis.
7. Use of adenosine monophosphate according to claim 1 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The medicine is used for inhibiting expression of inflammatory factor IL-6 protein.
8. Use of adenosine monophosphate according to claim 1 for the preparation of a medicament for the treatment of osteoarthritis, characterized in that, The medicine is used for inhibiting expression of matrix metalloproteinase MMP13 protein.