Synergistic carrier for treating intervertebral disc degeneration through metabolism-inflammation axis intervention

Abstract

The invention belongs to the field of biological medicine, discloses a synergistic carrier for treating intervertebral disc degeneration through metabolism-inflammation axis intervention and a preparation method and application thereof, and belongs to the field of biological medicine. The synergistic carrier is of a nanoscale metal-polyphenol network structure and comprises a network core formed by metal ions and kaempferol through coordination and a Pluronic F127 polymer layer covering the surface of the core. And the average particle size of the material is 100-150 nm. The preparation method comprises the following steps: reacting kaempferol with a metal ion solution according to a molar ratio of 2: 1 to form a complexing core, coating with Pluronic F127, and freeze-drying. The synergistic carrier is taken as a core, and a pharmaceutical composition containing a pharmaceutically acceptable carrier is prepared. The carrier can be used for preparing drugs for treating intervertebral disc degeneration, and especially can be administered through local injection. According to the invention, kaempferol is used as a drug and a ligand at the same time, a synergistic treatment system with one drug and double effects is constructed, and a new material is provided for etiological treatment of intervertebral disc degeneration.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, and specifically discloses a synergistic carrier for treating intervertebral disc degeneration through metabolic-inflammatory axis intervention. Background Technology

[0002] Intervertebral disc degeneration (IDD) is a major cause of chronic low back pain and spinal dysfunction, and its incidence continues to rise with population aging and the prevalence of sedentary lifestyles. Studies published in journals such as *The Lancet* indicate that the number of people suffering from low back pain has increased by more than 50% in the past thirty years, and IDD has become a disease that seriously affects public health and the workforce. The intervertebral disc is mainly composed of the nucleus pulposus, annulus fibrosus, and cartilaginous endplates, and its structural stability depends on the dynamic balance of the extracellular matrix (ECM). Long-term mechanical stress, inflammation, and metabolic disorders can lead to impaired nucleus pulposus cell function and decreased matrix synthesis, resulting in reduced disc height and nerve compression symptoms.

[0003] Current treatment options mainly include conservative therapy and surgery. Conservative treatments, such as nonsteroidal anti-inflammatory drugs (NSAIDs) and physical rehabilitation, can only provide short-term symptom relief and cannot reverse degeneration. While surgery can improve pain, it is highly invasive, expensive, has a high recurrence rate, and cannot block disease progression at the molecular level. Therefore, developing safe, effective, and etiologically sound new strategies to intervene in the degenerative process is an important direction for current research.

[0004] In recent years, multi-omics studies have revealed that intervertebral disc degeneration is not only structural deterioration but also involves a mutually reinforcing process of "metabolic disorder-inflammatory response." Key glycolytic enzymes such as ENO1, PKM, and TPI1 are significantly upregulated in degenerated tissues, leading to lactic acid accumulation and local acidification. This further activates the NF-κB pathway, inducing the continuous release of inflammatory factors (IL-1β, IL-6, MMPs, ADAMTSs), forming a "metabolic-inflammatory positive feedback loop." Targeting this mechanism, natural active substances such as kaempferol and shikonin have shown anti-inflammatory and metabolic regulatory effects at the cellular level. However, due to their high lipid solubility, poor water solubility, and rapid metabolism in vivo, they are difficult to achieve effective concentrations locally in the intervertebral disc, resulting in limited therapeutic effects.

[0005] In summary, the existing technologies have the following main shortcomings: First, the mechanism of action is singular, making it difficult to simultaneously regulate metabolic imbalance and inflammatory response; second, natural drugs have poor stability and low bioavailability; third, the safety of existing carrier systems needs to be improved; and fourth, there is a lack of synergistic effects and system validation of dual drugs. Summary of the Invention

[0006] To address the aforementioned problems, this invention discloses a synergistic carrier for treating intervertebral disc degeneration through metabolic-inflammatory axis intervention. This carrier uses kaempferol as both a therapeutic drug and a ligand, self-assembling with metal ions to form a metal-polyphenol network core, which is then encapsulated with Pluronic F127 to construct a nanoscale synergistic carrier. It can simultaneously inhibit inflammatory responses and correct metabolic disorders in the degenerated disc region, thereby precisely breaking the vicious cycle of "metabolic-inflammatory." This carrier possesses both good biocompatibility and synergistic therapeutic effects, providing a new material for the etiological treatment of intervertebral disc degeneration.

[0007] To achieve the objectives of this invention, the invention includes the following technical solutions:

[0008] A synergistic carrier for treating intervertebral disc degeneration through metabolic-inflammatory axis intervention, wherein the synergistic carrier is a nanoscale metal-polyphenol network structure, comprising:

[0009] 1) A metal-polyphenol network core formed by coordination between metal ions and polyphenol compounds, wherein the polyphenol compound is kaempferol;

[0010] 2) An amphiphilic block copolymer layer covering the surface of the core.

[0011] Furthermore, in the aforementioned synergistic carrier, the metal ion is Fe. 3+ .

[0012] Furthermore, in the aforementioned synergistic carrier, the amphiphilic block copolymer is Pluronic F127.

[0013] Furthermore, in the aforementioned synergistic carrier, the mass fraction of Pluronic F127 accounts for 20-30% of the core mass of the metal-polyphenol network.

[0014] Furthermore, the above-mentioned synergistic carrier has an average particle size of 100-150 nm.

[0015] This invention also discloses a method for preparing the above-mentioned synergistic carrier, comprising the following steps:

[0016] S1. Dissolve kaempferol in an ethanol-water mixed solvent to form a kaempferol solution;

[0017] S2. Under stirring conditions, a metal ion solution is added dropwise to the kaempferol solution, controlling the molar ratio of kaempferol to metal ions to be 2:1. The reaction is carried out for 2-4 hours to form a kaempferol-metal complex precipitate.

[0018] S3. Centrifuge and wash the precipitate to obtain the metal-polyphenol network core;

[0019] S4. The core is redispersed in a solvent, an amphiphilic block copolymer solution is added, and the mixture is stirred for 6-8 hours. After ultrasonic dispersion and centrifugation, the precipitate is collected and freeze-dried to obtain the synergistic carrier.

[0020] The present invention also discloses a pharmaceutical composition comprising any of the synergistic carriers described above and a pharmaceutically acceptable carrier or diluent.

[0021] The present invention also discloses the use of the above-mentioned synergistic carrier in the preparation of a drug for treating intervertebral disc degeneration.

[0022] Furthermore, in the above-mentioned uses, the drug is an injectable preparation, administered to the degenerated area of ​​the intervertebral disc via local injection.

[0023] Compared with the prior art, the present invention has the following outstanding advantages:

[0024] This invention innovatively constructs a drug delivery system based on a metal-polyphenol network, using kaempferol as both a therapeutic drug and a ligand, achieving synergistic treatment with "one drug, two effects," while also realizing the following technical advantages:

[0025] (1) Through the dual pharmacological effects of kaempferol's own metabolic regulation and anti-inflammatory properties, it can simultaneously intervene in metabolic abnormalities and inflammatory responses at the molecular level, thus blocking their vicious cycle;

[0026] (2) A simple and green "one-step coordination-surface coating" self-assembly process is adopted, combined with the biocompatible Pluronic F127 for modification. While ensuring the stability of the carrier and the drug loading rate, the biocompatibility and industrialization feasibility of the material are significantly improved. The invention has been verified by in vitro and in vivo experiments to effectively delay the process of intervertebral disc degeneration. Its mechanism of action is highly consistent with the key targets found in multi-omics studies, providing a new material platform and technical path for the precision treatment of intervertebral disc degeneration. Attached Figure Description

[0027] Figure 1 Tissue proteomics analysis reveals differentially expressed protein characteristics. A. Tissue proteomics flowchart; B. Patient MRI images; C. H&E staining of nucleus pulposus tissue; D. Safranin-O staining of nucleus pulposus tissue; E. Statistical analysis of DIA-specific proteins in histological proteomics of grade II and V patients (6 vs 10); F. PCA analysis results of grade II and V patients; G. Volcano plot of differentially expressed proteins in DIA proteomics; H. Statistical analysis of differentially expressed proteins; I. Heatmap of differentially expressed proteins.

[0028] Figure 2Tissue proteomics enrichment analysis and validation: A. GO enrichment analysis results of upregulated proteins; B. KEGG enrichment analysis results of upregulated proteins; C. PPI analysis of glycolysis pathway; D. PPI analysis of immune regulation pathway; E. Violin plot representing the expression levels of proteins related to glycolysis pathway; F. Violin plot representing the expression levels of proteins related to immune regulation pathway; G. Validation of differentially expressed proteins in hypoxia-induced cell models.

[0029] Figure 3 Characterization of nanocomposites: A. Scanning electron microscopy structural characterization of kaempferol-iron and kaempferol-iron@F127; B. Adhesion experiment of kaempferol-iron@F127 to six organs: lung, liver, spleen, heart, and spinal cord; C. Thermogravimetric analysis results of kaempferol-iron@F127; D. Particle size changes of kaempferol-iron and kaempferol-iron@F127 in an aqueous dispersion system for 7 days; E. Surface charge changes of kaempferol-iron and kaempferol-iron@F127 in an aqueous dispersion system for 7 days.

[0030] Figure 4 The study included: A) In vitro cell experiments to verify the effects of the drug on cell morphology; B) CCK-8 assay to detect the cell proliferation of hNPC cells under different treatment conditions; C) Western blotting experiments to verify the effects of free drug and composite carrier on the expression levels of disease biomarkers MMP3 and MMP13 proteins; and D) Western blotting experiments to verify the effects of the composite carrier on the expression levels of disease biomarkers MMP3 and MMP13, as well as the key glycolysis protein ENO1 and the key immunomodulatory protein IL-6.

[0031] Figure 5 Animal puncture model validation: A. Modeling flowchart; B. Postoperative mouse weight change; C. MRI imaging results; D. Tissue section staining results; E. Quantification of intervertebral disc signal intensity; F. Quantification of intervertebral disc sagittal area. Detailed Implementation

[0032] Specifically, the technical solution of the present invention includes the following main parts:

[0033] Construction of nanocomposite supports: utilizing metal ions (Fe) 3+The drug kaempferol (PF127) forms a stable metal-phenol network (MPN) structure through coordination with polyphenol ligands. Loading and self-assembly of the material: PF127 molecules are introduced simultaneously during network formation, allowing them to embed into the network through hydrophobic interactions and hydrogen bonds, forming a drug-loaded composite system. Material characterization and performance evaluation: Scanning electron microscopy (SEM) reveals that the carrier exhibits a regular spherical or near-spherical nanostructure. Zeta potential analysis characterizes its surface charge and colloidal stability, confirming that Pluronic F127 coating maintains long-term stable dispersion in aqueous solution. Thermogravimetric analysis (TGA) assesses the carrier's thermal stability, demonstrating its excellent thermal tolerance. Cell and animal experiments validate the therapeutic effects of this drug-loaded system in anti-inflammatory, metabolic regulation, and matrix repair promotion in an in vitro nucleus pulposus cell inflammation model and an in vivo mouse caudal puncture degeneration model.

[0034] (I) Construction of nanocomposite carriers

[0035] Raw material composition

[0036] 1. Metal ion source: Ferric chloride (FeCl3·6H2O, analytical grade); 2. Polyphenol ligand and drug molecule: Kaempferol (≥98%, Sigma-Aldrich); 3. Polymer carrier: Pluronic F127 (PEO) 100 –PPO 65 –PEO 100 (The average molecular weight is approximately 12,600); 4. Solvent system: Anhydrous ethanol and deionized water are mixed at a ratio of 1:1 (v / v).

[0037] Preparation of Kaempferol–Fe Complex

[0038] Kaempferol was dissolved in an ethanol-water mixture (final concentration 0.5–1.0 mg / mL), and FeCl3 solution (1–5 mM) was slowly added dropwise under magnetic stirring, controlling the molar ratio of Kaempferol:FeCl3. 3+ = 2:1. The reaction proceeds at room temperature for 2–4 hours, allowing Fe to... 3+ It coordinates with the hydroxyl groups of kaempferol to form a stable Kaempferol–Fe complex network structure.

[0039] The resulting solution was yellowish-brown, indicating successful MPN formation. Centrifugation (10,000 rpm, 10 min) removed unbound impurities, and the precipitate was collected and washed twice with deionized water.

[0040] F127 encapsulation modification (20–30 wt%)

[0041] The Kaempferol–Fe precipitate was redispersed in an ethanol-water (1:1) mixture, and a pre-dissolved Pluronic F127 solution (20–30 wt% relative to solid Kaempferol–Fe) was added. The mixture was magnetically stirred at room temperature for 6–8 h to allow F127 to be uniformly adsorbed onto the particle surface through hydrophobic-hydrophobic interactions and hydrogen bonding.

[0042] Subsequently, ultrasonic dispersion (80 W, 3 min) was used to promote the formation of a uniform coating layer. Free F127 was removed by centrifugation (8,000 rpm, 10 min), the precipitate was collected and lyophilized to obtain the Kaempferol–Fe@F127 nanocomposite.

[0043] 2. Storage and Characterization: The resulting lyophilized powder is light yellow and can be stored under drying conditions at −20°C.

[0044] (II) Cellular Experiment Verification

[0045] In a hypoxia-induced human nucleus pulposus (hNPCs) model, treatment with the vector system of this invention (Kaempferol-Fe@F127 nanocomposite carrier) did not significantly alter cell viability, but significantly decreased mRNA expression of inflammatory factors IL-1β, IL-6, and TNF-α, and upregulated matrix synthesis-related genes such as COL2A1 and Aggrecan. Western blot results showed decreased NF-κB p65 phosphorylation levels and downregulated expression of key glycolytic enzymes ENO1, PKM, and TPI1, suggesting that this system can achieve a dual effect of inflammation suppression and matrix repair by inhibiting metabolic reprogramming.

[0046] (III) Animal Experiment Verification

[0047] A disc degeneration model was established using C57BL / 6JGpt mice via caudal puncture. Two weeks after modeling, the vector complex of this invention was injected locally. MRI imaging analysis showed that the disc height index (DHI) in the treatment group was significantly higher than that in the control group, and the Pfirrmann grade was improved. Histological staining (HE, Safranin O) showed that the nucleus pulposus structure was well preserved, and inflammatory cell infiltration was reduced. Immunohistochemical results showed decreased MMP3 expression and increased COL2A1 and Aggrecan expression, indicating that this vector system can effectively inhibit the progression of degeneration.

[0048] The technical solutions of the present invention will be clearly and completely described below in the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0049] Unless otherwise specified, all reagents or instruments used in the embodiments of this invention are commercially available conventional reagent products.

[0050] Example 1

[0051] Therapeutic target discovery and mechanism elucidation based on histioproteomics

[0052] This invention utilizes quantitative tissue proteomics technology based on DIA acquisition mode, and the flowchart is shown in the figure ( Figure 1 A) A systematic analysis was performed on intervertebral disc tissue samples of different degeneration grades (Grade II and Grade V), with MRI analysis of patients at different grades. Figure 1 B), while simultaneously performing H&E on the patient's tissue sections. Figure 1 C) and Safranin-O ( Figure 1 D) Staining, DIA proteomics identified approximately 800-1500 specific proteins in different samples. Figure 1 E) A total of 197 target proteins with significant expression differences were identified, and PCA characterization showed a good grouping effect. Figure 1 F), and a volcano plot was generated to show the number of differentially expressed proteins (F). Figure 1 G) and the representation of statistical graphs ( Figure 1 H), heatmap analysis was used to analyze the expression patterns of differentially expressed proteins ( Figure 1 I); For upregulated differentially expressed proteins, GO ( Figure 2 A) and KEGG ( Figure 2 B) Pathway enrichment analysis showed that the key enzymes in the glycolysis pathway, TPI1, PKM, PKLR, PGAM1, PGK1, and ENO1, were consistently upregulated in severely degenerated tissues. Figure 2 C); Simultaneously, inflammation-related proteins C3 and C5 were found to be significantly upregulated (C); Figure 2 D), the quantification of relevant proteins by proteomics is characterized by violin diagrams ( Figure 2E and F), their trends were positively correlated with the expression levels of metabolic enzymes, confirming a positive feedback loop between metabolic abnormalities and inflammatory responses at the molecular level. Simultaneously, we constructed a hypoxia-induced human nucleus pulposus (hNPC) intervertebral disc degeneration model and validated differentially expressed proteins identified through omics screening using Western blotting experiments. We found that this cell model exhibited a similar differential protein expression pattern to that of patients. Figure 2 G). These tissue proteomics findings directly reveal the core driving role of the "metabolic-inflammatory axis" in the process of intervertebral disc degeneration, providing key targets and theoretical basis for the therapeutic strategy of synergistic intervention of the glycolysis and inflammation pathways in this invention.

[0053] Example 2

[0054] Preparation and characterization of metal-polyphenol network dual drug carriers

[0055] Preparation principle and steps: using Fe 3+ The metal center coordinates with the ortho-phenolic hydroxyl groups in the kaempferol molecule, self-assembling to form a metal-polyphenol network (MPN) framework. This kaempferol simultaneously serves as both the active pharmaceutical ingredient and a ligand, directly participating in the construction of the drug-carrying network. Subsequently, through hydrophobic interactions and hydrogen bonding, the amphiphilic block copolymer Pluronic F127 was introduced to coat the surface of the aforementioned Kaempferol-Fe particles. Finally, after centrifugation purification and lyophilization, a structurally stable Kaempferol-Fe@F127 nanocomposite carrier was obtained.

[0056] The specific steps are as follows:

[0057] 1. Raw material preparation

[0058] 1) Source of metal ions: Ferric chloride (FeCl3·6H2O, analytical grade); 2) Phenolic ligand and drug molecule: Kaempferol (≥98%, Sigma-Aldrich); 3) Polymer carrier: Pluronic F127 (PEO) 100 –PPO 65 –PEO 100 (The average molecular weight is about 12,600); 4) Solvent system: Anhydrous ethanol and deionized water are mixed at a ratio of 1:1 (v / v).

[0059] 2. Preparation of Kaempferol-Fe Complex

[0060] Kaempferol was dissolved in an ethanol-water mixture (final concentration 0.75 mg / mL), and FeCl3 solution (3 mM) was slowly added dropwise under magnetic stirring, controlling the molar ratio of Kaempferol:FeCl3. 3+= 2:1. The reaction proceeded at room temperature for 3 h, allowing Fe to... 3+ It coordinates with the hydroxyl groups of kaempferol to form a stable Kaempferol–Fe complex network structure.

[0061] The resulting solution was yellowish-brown, indicating successful MPN formation. Centrifugation (10,000 rpm, 10 min) removed unbound impurities, and the precipitate was collected and washed twice with deionized water.

[0062] 3. F127 coating modification (25wt%)

[0063] The Kaempferol–Fe precipitate was redispersed in an ethanol-water (1:1) mixture, and a pre-dissolved Pluronic F127 solution (25 wt% relative to solid Kaempferol–Fe) was added. The mixture was magnetically stirred at room temperature for 7 h to allow F127 to be uniformly adsorbed onto the particle surface through hydrophobic-hydrophobic interactions and hydrogen bonding.

[0064] 4. Subsequently, ultrasonic dispersion (80 W, 3 min) was used to promote the uniform formation of the coating layer. Free F127 was removed by centrifugation (8,000 rpm, 10 min), the precipitate was collected and freeze-dried to obtain Kaempferol–Fe@F127 nanocomposite.

[0065] Structural characteristics and performance description: Under scanning electron microscopy (SEM), this composite material exhibits a regular spherical or near-spherical nanostructure with uniform particle size distribution, averaging 100–150 nm. It also demonstrates good redispersibility and colloidal stability in water. Figure 3 A). Organ adhesion experiments demonstrated that this structure possesses good adhesion. Figure 3 B). Furthermore, thermogravimetric analysis (TGA) demonstrated that this nanocomposite support exhibited excellent thermal stability, maintaining significant mass retention even at 800°C. Figure 3 (C) This indicates that the constructed MPN and the coated Pluronic F127 possess high thermal stability, meeting the requirements of relevant application scenarios. Its surface carries a negative charge, which helps promote cellular uptake. Long-term tracking of the hydration size and Zeta potential of the nanoparticles further confirmed that the composite support coated with Pluronic F127 exhibited similar particle size distribution (Figure 3D) and surface charge (C) during a 7-day aqueous dispersion. Figure 3 E) No significant changes were observed, demonstrating significantly better colloidal stability than the uncoated core, which provides important assurance for the actual storage and in vivo application of the formulation.

[0066] Example 3

[0067] In vitro cell experiment design and preliminary validation

[0068] (1) Cell culture: Human nucleus pulposus cells (hNPCs) were cultured in a special medium (product number: IM-H497-1, Yimo Biotechnology) and placed in a constant temperature incubator at 37℃ and 5% CO2.

[0069] Culture the cells. When the cells reach 80-90% confluence, digest and passage them using 0.25% trypsin.

[0070] (2) Establishment of a cell model for intervertebral disc degeneration: To simulate the pathological environment of intervertebral disc degeneration, a hypoxic culture method was used to construct a cell model. hNPCs were cultured at 1×10⁻⁶ cells per well. 5 Cells were seeded at a density of 1000 cells per cell in a culture plate. After the cells adhered, the culture plate was transferred to a three-gas incubator and cultured for 24 hours under conditions of 1% O2, 5% CO2 and 94% N2 to induce cell degeneration.

[0071] (3) Experimental grouping and treatment: The following experimental groups were set up: a. Control group: cells cultured under normal oxygen conditions; b. Model group: cells cultured under hypoxic conditions; Based on the two groups a and b, solvent, KPF-Fe and KPF-Fe@F127 were added to each group to construct the solvent group, KPF-Fe group and KPF-Fe@F127 group to explore the effects of the composite carrier drug on cells under normal conditions and hypoxic conditions. In the end, control-solvent, control-KPF-Fe, control-KPF-Fe@F127, hypoxia-solvent, hypoxia-KPF-Fe and hypoxia-KPF-Fe@F127 were constructed, for a total of 6 groups.

[0072] (4) Detection methods and procedures:

[0073] a. Cell viability assay: The CCK-8 assay was used. After treatment, 10% volume of CCK-8 solution was added to each well, and the cells were cultured for another 2 hours. The absorbance of each well was then measured at 450 nm using a microplate reader.

[0074] b. Western Blot: Total cellular protein was extracted using RIPA lysis buffer, and protein concentration was determined by the BCA method. Equal volumes of protein were subjected to SDS-PAGE electrophoresis and then transferred to a PVDF membrane. After blocking with 5% skim milk, the membrane was incubated overnight at 4°C with primary antibodies against ENO1, MMP-3, and the internal control β-actin, respectively. The following day, the membrane was incubated at room temperature for 1 hour with the corresponding peroxidase-labeled secondary antibody. Finally, the membrane was developed using an ECL chemiluminescence assay kit, and the data were analyzed.

[0075] Based on preliminary cell experiments, the material showed no significant toxicity to hNPCs cells, with cell viability >90%. Figure 4 A). CCK-8 assays showed that the vector groups were essentially indistinguishable under normal culture conditions (con), but significantly enhanced cell viability under hypoxic conditions (HX), indicating that the vector has good biocompatibility. Figure 4 B). Simultaneously, Western blot results confirmed that the vector group could regulate the expression of the NF-κB inflammatory pathway (IL-6) and the key glycolytic protein (ENO1). These results collectively demonstrate that the composite vector of this invention possesses synergistic anti-inflammatory and metabolic regulatory functions. Figure 4 C, D).

[0076] Example 4

[0077] Animal models and validation pathways

[0078] (1) Experimental animals: Forty male C57BL / 6JGpt mice aged 6-8 weeks and weighing 20-27g were used in this study.

[0079] (2) Establishment of the intervertebral disc degeneration model: The coccygeal puncture method was used. Mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital and fixed in a prone position. The skin of the tail was disinfected, and a longitudinal incision was made in the midline of the Co6 / 7 and Co7 / 8 segments of the coccyx (corresponding to the two segments in the figure) to expose the annulus fibrosus (the two intervertebral disc segments Co6 / 7 of the coccyx were punctured, and Co5 / 6 was exposed as an in vivo control). A 27G needle (about 4 mm long) was used to puncture the nucleus pulposus perpendicular to the surface of the annulus fibrosus and rotated 360° for 1 min to cause controlled intervertebral disc damage.

[0080] (3) Grouping and administration: Mice were randomly divided into 4 groups (n=10): a. Sham group (only the skin was cut and the tailbone was exposed, without puncture); b. Puncture group (puncture + injection of an equal volume of PBS); c. Free drug group (puncture + injection of about 50 μg of free KPF-Fe drug); d. Carrier treatment group (puncture + injection of about 50 μg of KPF-Fe@F127 composite carrier).

[0081] Animal characterization analysis was performed 4 weeks after the animal experiment. Figure 5 A). Four weeks post-surgery, there were no significant differences in body weight among all experimental mice. Figure 5B). During modeling, the composite carrier and free drug of this invention were locally injected, and the intervertebral disc height index (DHI) and Pfirrmann grade were assessed by MRI. Results showed that the carrier treatment group (corresponding to "Puncture & KPF-Fe@F127" in the figure) had significantly improved signal compared to the puncture group ("Puncture"), and the Pfirrmann grade was superior to the control group and the free drug group. Figure 5 C). Meanwhile, both mouse liver and tailbone sections showed that the KPF-Fe@F127 composite carrier drug group did not exhibit significant liver morphological changes compared to other groups, and the tailbone intervertebral disc structure also maintained a relatively intact morphology. Figure 5 D). Analysis of imaging results showed that the KPF-Fe@F127 mice in the composite carrier drug group had good signal intensity in the intervertebral discs (D). Figure 5 E), the sagittal section area of ​​the intervertebral disc in MRI maintained a good level ( Figure 5 F).

[0082] In summary, preliminary results from animal experiments demonstrate that the material of this invention can effectively slow down the degenerative process of the intervertebral disc. The groups shown in the images intuitively illustrate the superiority of carrier therapy.

[0083] Security and Application Prospects

[0084] The components of the metal-polyphenol network are all biocompatible, and their degradation products can be metabolized and absorbed without causing long-term accumulation. This material system can be further developed into injectable sustained-release formulations or hydrogels, suitable for precise local drug delivery in clinical settings.

[0085] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the above embodiments do not limit the present invention in any way, and all technical solutions obtained by equivalent substitution or equivalent transformation fall within the protection scope of the present invention.

Claims

1. A synergistic carrier for treating intervertebral disc degeneration through metabolic-inflammatory axis intervention, characterized in that, The synergistic carrier is a nanoscale metal-polyphenol network structure, comprising: 1) A metal-polyphenol network core formed by coordination between metal ions and polyphenol compounds, wherein the polyphenol compound is kaempferol; 2) An amphiphilic block copolymer layer covering the surface of the core.

2. The collaborative carrier according to claim 1, characterized in that, The metal ion is Fe. 3+ .

3. The collaborative carrier according to claim 1, characterized in that, The amphiphilic block copolymer is Pluronic F127.

4. The collaborative carrier according to claim 3, characterized in that, The mass fraction of Pluronic F127 accounts for 20-30% of the core mass of the metal-polyphenol network.

5. The collaborative carrier according to claim 1, characterized in that, The average particle size of the synergistic carrier is 100-150 nm.

6. The method for preparing the synergistic carrier according to any one of claims 1-5, characterized in that, Includes the following steps: S1. Dissolve kaempferol in an ethanol-water mixed solvent to form a kaempferol solution; S2. Under stirring conditions, add metal ion solution dropwise to the kaempferol solution, control the molar ratio of kaempferol to metal ions to be 2:1, react for 2-4 hours, and form kaempferol-metal complex precipitate. S3. Centrifuge and wash the precipitate to obtain the metal-polyphenol network core; S4. The core is redispersed in a solvent, an amphiphilic block copolymer solution is added, and the mixture is stirred for 6-8 hours. After ultrasonic dispersion and centrifugation, the precipitate is collected and freeze-dried to obtain the synergistic carrier.

7. A pharmaceutical composition, characterized in that, It includes the synergistic carrier as described in any one of claims 1-5, as well as a pharmaceutically acceptable carrier or diluent.

8. Use of the synergistic carrier according to any one of claims 1-5 in the preparation of a medicament for treating intervertebral disc degeneration.

9. The use according to claim 8, characterized in that, The drug is an injectable preparation, administered via local injection to the degenerated area of ​​the intervertebral disc.

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