Application of BRD-K20733377 in preparation of medicine for treating intervertebral disc degeneration
By preparing BRD-K20733377 into an injection form and directly injecting it into the body, the problem of reduced number and functional decline of endogenous MSCs was solved, intervertebral disc degeneration was significantly improved, and effective therapeutic effects were achieved.
Patent Information
- Application Number
- CN202510877199.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-27
- Publication Date
- 2025-09-09
AI Technical Summary
Existing technologies are difficult to effectively repair and reconstruct the structure and function of degenerated intervertebral discs. The reduction in the number and function of endogenous MSCs leads to insufficient endogenous repair effects, and there is a lack of effective therapeutic drugs for intervertebral disc degeneration.
BRD-K20733377 is used as the active ingredient and prepared into an injection form, which is directly injected into the body. It has the function of resisting ferroptosis of MSCs, especially NPMSCs, alleviating erastin-induced damage, improving extracellular matrix degradation, and delaying intervertebral disc degeneration.
BRD-K20733377 significantly improves NPMSCs ferroptosis, reduces extracellular matrix degradation, and delays intervertebral disc degeneration. It has significant effects and a low dosage, which is superior to oral Western medicine.
Smart Images
Figure CN120605271A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of medical technology, and in particular to an application of BRD-K20733377 in preparing a drug for treating intervertebral disc degeneration. Background Art
[0002] Intervertebral disc degeneration (IVDD) is a major pathological factor causing cervical, lumbar, and back pain, and plays a significant role in the world's major health problems. IVDD is generally considered one of the primary initiating factors for degenerative disc diseases, including cervical, shoulder, lumbar, and leg pain. Currently, conventional treatments for IVDD (conservative or surgical) are symptomatic and fail to repair and reconstruct the structure and function of the degenerated disc. Biological therapies based on mesenchymal stem cell (MSC) transplantation show promise in treating IVDD, but the long-term survival of transplanted MSCs remains a major obstacle to current MSC-based biological therapies. In recent years, endogenous MSC-driven repair has garnered increasing attention. The unfavorable local microenvironment of the degenerated disc leads to a decrease in the number and function of endogenous nucleus pulposus-derived mesenchymal stem cells (NPMSCs), preventing them from effectively exerting their endogenous repair functions. This may be another major cause of IVDD. Currently, there is no clinical treatment for IVDD. It is of great significance to discover new drugs that can effectively treat IVDD.
[0003] In view of this, the present invention is proposed. Summary of the Invention
[0004] In order to solve the above technical problems, the present invention provides a use of BRD-K20733377 in the preparation of a drug for treating intervertebral disc degeneration.
[0005] Specifically, the technical solution of the present invention is as follows:
[0006] In a first aspect, the present invention provides the use of BRD-K20733377 in the preparation of a drug for treating intervertebral disc degeneration. The structural formula of the BRD-K20733377 is shown in formula (I):
[0007]
[0008] Preferably, the drug for treating intervertebral disc degeneration has the function of resisting MSCs ferroptosis.
[0009] Preferably, the MSCs are NPMSCs.
[0010] Preferably, the drug for treating intervertebral disc degeneration has the function of reducing the damage caused by ferroptosis inducers to MSCs.
[0011] Preferably, the ferroptosis inducer comprises Erastin.
[0012] Preferably, the dosage form of the drug for treating intervertebral disc degeneration is an injection.
[0013] In a second aspect, the present invention provides a drug for treating intervertebral disc degeneration, wherein the active ingredient of the drug includes BRD-K20733377.
[0014] Preferably, the dosage form of the drug is an injection.
[0015] Preferably, in the drug, in addition to BRD-K20733377, the active ingredient of the drug further includes MSCs.
[0016] As another preferred embodiment, in the drug, BRD-K20733377 is the only active ingredient of the drug; the drug is used for the treatment of intervertebral disc degeneration in combination with MSCs.
[0017] Beneficial effects:
[0018] The present invention provides an application of BRD-K20733377 in the preparation of a drug for treating intervertebral disc degeneration. The present invention's research found that BRD-K20733377 has an anti-ferroptosis effect on NPMSCs and can improve Erastin-induced ferroptosis in NPMSCs. In addition, in vivo experiments on rat animal models showed that BRD-K20733377 can reduce the degradation of the extracellular matrix of NPMSCs and delay intervertebral disc degeneration. The present invention provides an application of BRD-K20733377 in the preparation of a drug for treating intervertebral disc degeneration. Compared with oral Western medicine, BRD-K20733377 can be administered by direct injection, and only a very small amount of injection is required to achieve the therapeutic purpose, with significant effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] In order to more clearly illustrate the technical solutions of the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be described below.
[0020] Figure 1 : Morphological photos of NPMSCs. Primary NPMSCs are slender, spindle-shaped and grow in a sunflower-like pattern.
[0021] Figure 2: Cell viability and cell proliferation assay results. NPMSCs were treated with Cell Counting Kit-8 (CCK-8), different concentrations of BRD-K20733377 and Erastin.
[0022] Figure 3 : Cell cycle assay results. Flow cytometry cell cycle assay of NPMSCs in different groups.
[0023] Figure 4 :Reactive oxygen species (ROS) fluorescence detection results of NPMSCs. The expression levels of ROS in different groups were detected by immunofluorescence.
[0024] Figure 5 Mitochondrial membrane potential (MMP) detection results. The fluorescence intensity in different groups was measured using the JC-1 detection kit.
[0025] Figure 6 :FerroOrange staining results. 2+ content.
[0026] Figure 7 :Liperfluo test results. Detection of lipid peroxide levels in different groups.
[0027] Figure 8 :Transmission electron microscopy images of mitochondria. The morphology of mitochondria in different groups of cells was observed.
[0028] Figure 9 :Malondialdehyde test results. The amount of malondialdehyde in different groups was tested.
[0029] Figure 10 : Western blot analysis results of ferroptosis-related proteins. Quantitative analysis of the expression of FTH1, GPX4, SLC7A11, and ACSL4 in different groups.
[0030] Figure 11 : Results of immunofluorescence analysis of ferroptosis-related proteins. Quantitative analysis of the expression of FTH1, GPX4, SLC7A11, and ACSL4 in different groups.
[0031] Figure 12 : X-ray examination (DHI quantification) and MRI examination (thermal map quantification) results of rat caudal vertebrae and photos of histological specimens.
[0032] Figure 13 : Photos of the results of Safranin O-Fast Green staining, hematoxylin and eosin staining, immunohistochemistry of type II collagen (COL2), and immunohistochemistry of matrix metalloproteinase 13 (MMP13) of rat intervertebral disc.
[0033] Figure 14: Molecular structure of BRD-K20733377. DETAILED DESCRIPTION
[0034] The present invention aims to provide an application of BRD-K20733377 in alleviating damage caused by Erastin to NPMSCs and preventing ferroptosis of NPMSCs, and particularly relates to an application of BRD-K20733377 in delaying intervertebral disc degeneration.
[0035] The molecular formula of BRD-K20733377 is: C 33 H 40 O 19 , molecular weight: 740.66, its specific molecular structure can be found in Figure 14 .
[0036] BRD-K20733377 is considered to be a compound with anti-aging effects. Studies have found that BRD-K20733377 significantly reduces the load of senescent cells and the mRNA expression of aging-related genes in the kidneys of elderly mice. Ferroptosis is one of the important causes of intervertebral disc degeneration. However, the protective effect of BRD-K20733377 on NPMSCs is still unclear. Previous studies of the present invention have shown that BRD-K20733377 has common intersection targets with ferroptosis and IVDD, and further studied whether BRD-K20733377 can alleviate Erastin-induced ferroptosis of NPMSCs, and verified the effect of BRD-K20733377 in alleviating intervertebral disc degeneration in in vitro and in vivo experiments.
[0037] The research results of the present invention show that BRD-K20733377 has the effects of resisting ferroptosis of NPMSCs and delaying intervertebral disc degeneration.
[0038] Based on the above research results, the present invention further proposes a pharmaceutical preparation for treating and delaying intervertebral disc degeneration, wherein the active ingredient includes BRD-K20733377. Furthermore, the dosage form of the drug includes an injection.
[0039] The beneficial effects of the present invention are primarily manifested in: BRD-K20733377 exhibits an anti-ferroptosis effect on NPMSCs, ameliorating erastin-induced ferroptosis in NPMSCs. In vivo experiments in rat models have demonstrated that BRD-K20733377 can reduce the degradation of the extracellular matrix of NPMSCs and delay intervertebral disc degeneration. Therefore, BRD-K20733377 has significant potential in the treatment of degenerative disc diseases. Furthermore, compared to oral Western medications, direct injection is more effective, requiring only a minimal injection to achieve therapeutic effects.
[0040] The following examples provide a detailed description of the technical solutions provided by the present invention, but they should not be construed as limiting the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the examples are conventional methods; the materials and reagents used are all commercially available.
[0041] Example 1
[0042] This example provides the following experimental content to demonstrate that BRD-K20733377 has the effect of inhibiting ferroptosis of NPMSCs and delaying intervertebral disc degeneration.
[0043] (1) Characteristics of NPMSCs.
[0044] Spindle-shaped NPMSCs were isolated and cultured from rat intervertebral discs. Figure 1 .
[0045] (2) Effect of BRD-K20733377 on NPMSCs cell viability and proliferation ability.
[0046] The CCK-8 assay was used to evaluate the effects of BRD-K20733377 and Erastin on the viability of NPMSCs.
[0047] The results are as follows Figure 2 As shown, BRD-K20733377 had no obvious cytotoxic effect at a concentration of 6 μM for 72 hours.
[0048] based on Figure 2 As a result, BRD-K20733377 was selected at a concentration of 6 μM in the following experiments. After Erastin treatment, cell viability decreased in a dose-dependent manner, and then the cells were treated with 5 μM Erastin for the experiment.
[0049] (3) Effect of BRD-K20733377 on the cell cycle of NPMSCs.
[0050] Flow cytometry analysis of cell cycle is one of the markers for evaluating cell senescence.
[0051] The results are as follows Figure 3 As shown, the G2 / M phase ratio of NPMSCs in the BRD-K20733377 treatment group was significantly higher than that in the Erastin group.
[0052] (4) BRD-K20733377 inhibits Erastin-induced ferroptosis of NPMSCs.
[0053] To explore the protective effect of BRD-K20733377 on Erastin-induced ferroptosis in NPMSCs. This example uses reactive oxygen species detection, mitochondrial membrane potential detection, FerroOrange detection, Liperfluo detection, mitochondrial transmission electron microscopy, malondialdehyde detection, Western blot, and immunofluorescence staining. Figure 4-11 .
[0054] Figure 4 The results showed that BRD-K20733377 treatment could reduce the level of reactive oxygen species induced by erastin.
[0055] Figure 5 The results showed that BRD-K20733377 treatment could reduce the mitochondrial membrane potential level induced by Erastin.
[0056] Figure 6 The results showed that BRD-K20733377 treatment could reduce the erastin-induced Fe 2+ level.
[0057] Figure 7 The results showed that BRD-K20733377 treatment could reduce the level of lipid peroxides induced by Erastin.
[0058] Figure 8 The results showed that BRD-K20733377 treatment could reduce the disorder of mitochondrial cristae induced by erastin.
[0059] Figure 9 The results showed that BRD-K20733377 treatment could reduce erastin-induced malondialdehyde levels.
[0060] Figure 10 The results showed that BRD-K20733377 treatment could reduce the expression of erastin-induced ACSL4 and increase the expression of GPX4, FTH1 and SLC7A11.
[0061] Figure 11 The immunofluorescence results were consistent with those of Western blot.
[0062] In summary, the experimental results show that BRD-K20733377 plays a protective role in Erastin-induced ferroptosis of NPMSCs.
[0063] (5) BRD-K20733377 improves horizontal intervertebral disc degeneration in SD rat animal models.
[0064] In the PBS-injected group, DHI decreased significantly, while BRD-K20733377 treatment reversed this decrease. In the PBS-injected group, NP signal intensity decreased significantly and Pfirrmann grade increased significantly, while the BRD-treated group (treated with BRD-K20733377) reversed these changes. The nucleus pulposus area and histological scores of rat intervertebral disc specimens from different groups were further measured. In the PBS-injected group, the relative area of NP gradually decreased over time, while the histological score gradually increased; however, the BRD-treated group reversed these changes ( Figure 12 ). Rat intervertebral disc sections were stained using hematoxylin-eosin (HE) and safranin O / fast green (SF) staining techniques. Four weeks after PBS treatment, compression-induced deformation of the nucleus pulposus (NP) was observed, resulting in a reduction in its area of approximately 20-25%. The NP in the PBS-treated group almost completely disappeared, and the intervertebral space was replaced by chondrocytes extending inward. In contrast, the BRD-treated group significantly slowed the progression of IVDD. After SF staining, the NP tissue appeared red due to collagen accumulation, while the annulus fibrosus (AF) tissue appeared blue. In the PBS-treated group, the NP tissue area ratio was the lowest among all groups, and the damaged NP tissue was replaced by the blue annulus fibrosus. In contrast, the larger red area in the BRD-treated group still existed, indicating collagen enrichment, suggesting a better treatment effect. MMP13 immunohistochemistry results: In the control group, the NP appeared light brown; compared with the control group, the NP area in the PBS-treated group was reduced, and the histone staining intensity increased; compared with the PBS-treated group, the brown staining in the BRD-treated group was reduced, indicating downregulation of MMP13. The results of COL-II immunohistochemistry showed that the brown staining was the strongest in the control group. In contrast, the brown staining was the lightest in the PBS treatment group. The staining intensity in the BRD treatment group was higher than that in the PBS treatment group ( Figure 13 ).
[0065] The above results indicate that BRD-K20733377 can alleviate the progression of IVDD in rats induced by puncture.
[0066] The above-described embodiments merely illustrate several implementation methods of the present invention, and are provided to facilitate a specific and detailed understanding of the technical solutions of the present invention. They should not be construed as limiting the scope of the invention patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the scope of the present invention, and all such variations and improvements fall within the scope of protection of the present invention.
Claims
1. Use of BRD-K20733377 in the preparation of a drug for treating intervertebral disc degeneration, wherein the structural formula of BRD-K20733377 is shown in formula (I):
2. The application according to claim 1, characterized in that The drug for treating intervertebral disc degeneration has the function of resisting MSCs ferroptosis.
3. The application according to claim 2, characterized in that: The MSCs are NPMSCs.
4. The application according to claim 1, characterized in that The drug for treating intervertebral disc degeneration has the function of reducing the damage caused by ferroptosis inducers to MSCs.
5. The application according to claim 4, characterized in that: The ferroptosis inducer includes Erastin.
6. The use according to any one of claims 1 to 5, characterized in that: The dosage form of the drug for treating intervertebral disc degeneration is an injection.
7. A drug for treating intervertebral disc degeneration, characterized in that: The active ingredient of the drug includes BRD-K20733377.
8. The drug for treating intervertebral disc degeneration according to claim 7, characterized in that: The dosage form of the medicine is injection.
9. The drug for treating intervertebral disc degeneration according to claim 7 or 8, characterized in that: In addition to BRD-K20733377, the active ingredient of the drug also includes MSCs.
10. The drug for treating intervertebral disc degeneration according to claim 7 or 8, characterized in that: BRD-K20733377 is the sole active ingredient of the drug; the drug is used in combination with MSCs to treat intervertebral disc degeneration.