Novel Anti-in-stent-restenosis protein-eluting stent

Abstract

Disclosed in the present invention is a novel anti-in-stent-restenosis protein-eluting stent. Specifically, disclosed in the present invention is a medical stent. The stent comprises: (a) a supporting structure, wherein the surface of the supporting structure comprises a coating, and the coating contains polydopamine; and (b) a CCN5 recombinant protein, wherein the recombinant protein is immobilized to the supporting structure by binding to the polydopamine. The medical stent of the present invention is used for preventing in-stent restenosis after PCI, and is also used for promoting the proliferation and migration of a human aortic endothelial cell (HAEC) line and / or promoting endothelialization to inhibit in-stent restenosis.

Description

A new protein-coated stent to prevent in-stent restenosis Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a novel anti-in-stent restenosis protein-coated stent. Background Art

[0002] Myocardial infarction (MI) caused by atherosclerotic coronary heart disease (CAD) is a major threat to human health worldwide and ranks as the leading cause of death. Percutaneous coronary intervention (PCI) and coronary revascularization have become the primary treatment for CAD MI. Post-PCI endothelial cell loss and damage induce abnormal proliferation and migration of vascular smooth muscle cells (VSMCs), forming a neointima, leading to vascular restenosis and a major cause of long-term recurrent myocardial ischemia in patients after PCI stent placement.

[0003] Currently, there are several solutions for in-stent restenosis after PCI: drug-eluting stents (DES), which are coated with a broad-spectrum anti-cell proliferation drug that releases drugs to inhibit restenosis. Commonly used drugs include sirolimus and rapamycin; stent expansion, which involves performing another balloon angioplasty to widen the narrowed area within the stent to restore vascular patency; and placement of a new stent in the restenotic area to reopen coronary artery blood flow.

[0004] Currently, the drugs on the surface of drug-coated stents have a broad-spectrum inhibitory effect. While inhibiting the proliferation of smooth muscle cells, they also inhibit the repair of vascular endothelium, resulting in poor long-term in-stent endothelial repair after PCI and increasing the risk of long-term in-stent restenosis after PCI.

[0005] Balloon dilatation or re-implantation of a stent requires patients to undergo a second invasive treatment, which increases costs and does not address the risk of recurrence of in-stent restenosis in the long term after the second treatment. The present invention only requires a single treatment, reducing the possibility of patients needing repeated treatment.

[0006] Therefore, there is an urgent need in this field to develop new methods that can inhibit smooth muscle proliferation while promoting endothelial repair and reduce long-term in-stent restenosis after PCI. Summary of the Invention

[0007] The purpose of the present invention is to provide a new method that can inhibit smooth muscle proliferation while promoting endothelial repair, thereby reducing long-term in-stent restenosis after PCI surgery.

[0008] A first aspect of the present invention provides a medical stent, comprising:

[0009] (a) a supporting structure, wherein a surface of the supporting structure comprises a coating, wherein the coating comprises polydopamine;

[0010] (b) CCN5 recombinant protein, wherein the recombinant protein is fixed to the support structure by binding to polydopamine.

[0011] In another preferred embodiment, the recombinant protein is covalently bound to polydopamine and fixed to the support structure.

[0012] In another preferred embodiment, the support structure is selected from the group consisting of a coronary artery stent, a carotid artery stent, an abdominal aorta stent, a femoral artery stent, a renal artery stent, a pulmonary artery stent, a cerebral artery stent, or a combination thereof.

[0013] In another preferred embodiment, the quinone group of the polydopamine in the coating undergoes Michael addition reaction with the amino group of the CCN5 recombinant protein under alkaline conditions, thereby coating the CCN5 recombinant protein on the support structure.

[0014] In another preferred embodiment, the sequence of the CCN5 recombinant protein is shown as SEQ ID NO.1.

[0015] In another preferred embodiment, the loading amount of the CCN5 recombinant protein is 150-1000 ng / cm 2 , preferably, 250-800ng / cm 2 , more preferably, 300-500 ng / cm 2 .

[0016] In another preferred embodiment, the number of layers of polydopamine coated on the support structure is 1-6 layers, preferably 2-5 layers; more preferably 3-4 layers.

[0017] In another preferred embodiment, the weight ratio of the polydopamine and the CCN5 recombinant protein in the medical stent is 0.1%-2%, more preferably, 0.2%-1.5%, and most preferably, 0.5%-1%.

[0018] In another preferred embodiment, the material of the medical stent is selected from the following group: metal and non-metal.

[0019] In another preferred embodiment, the metal includes: a stainless steel stent, a titanium alloy stent, and a chromium-cobalt alloy stent.

[0020] In another preferred embodiment, the non-metallic material includes: a polylactic acid stent and a polyhydroxybutyric acid stent.

[0021] In another preferred embodiment, the shape of the medical stent is selected from the following group: circle, ellipse, or a combination thereof.

[0022] A second aspect of the present invention provides a method for manufacturing a medical stent, comprising the steps of:

[0023] (a) providing a support structure and CCN5 recombinant protein, wherein the surface of the support structure comprises a coating comprising polydopamine;

[0024] (b) incubating the support structure and the CCN5 recombinant protein under alkaline conditions, so that the quinone group of the polydopamine in the coating undergoes a Michael addition reaction with the amino group of the CCN5 recombinant protein under alkaline conditions, thereby coating the CCN5 recombinant protein on the support structure to obtain the medical stent.

[0025] In another preferred embodiment, the alkaline condition refers to a pH of 7.5-9.5, preferably 7.5-8.5.

[0026] In another preferred embodiment, the incubation time is 2-8 hours, preferably 3-6 hours.

[0027] The third aspect of the present invention provides a use of the medical stent described in the first aspect of the present invention for preparing a medical device for preventing in-stent restenosis after intravascular interventional treatment.

[0028] In another preferred embodiment, the medical device is further used for one or more purposes selected from the following groups:

[0029] (a) Promotes the proliferation and migration of human aortic endothelial cells (HAECs);

[0030] (b) Promote endothelialization and inhibit in-stent restenosis.

[0031] In another preferred embodiment, the intravascular interventional treatment includes PCI, neurovascular stent implantation, and peripheral vascular stent implantation.

[0032] In another preferred embodiment, the peripheral vascular stent implantation is selected from the group consisting of renal artery stent implantation, lower limb artery stent implantation, carotid artery stent implantation, or a combination thereof.

[0033] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1: CCN5 recombinant protein was successfully immobilized on the surface of a metal stent or circular metal sheet. A: X-ray photoelectron spectroscopy (XPS) confirmed the successful immobilization of CCN5 recombinant protein on the surface of a metal stent or circular metal sheet. B: Quartz crystal microbalance showed that the loading amount of CCN5 recombinant protein on the stent metal material surface was approximately 330 ng / cm 2 .

[0035] Figure 2: Using plain circular metal sheets, circular metal sheets coated with polydopamine (PDA), or circular metal sheets coated with bovine serum albumin (BSA) as controls, in vitro experiments show that circular metal sheets coated with CCN5 recombinant protein promote the adhesion, proliferation, and migration of human aortic endothelial cells (HAECs). A: HAECs were plated on the surfaces of each circular metal sheet and adhered HAECs were observed after 2 hours, demonstrating that CCN5 recombinant protein coating promoted HAEC adhesion. BC: HAECs were plated on the surfaces of each circular metal sheet, and Ki67 staining (B) and MTT assay (C) confirmed that CCN5 recombinant protein coating significantly promoted HAEC proliferation. D: HAECs were fully plated on the surfaces of each circular metal sheet, and a scratch assay revealed that CCN5 recombinant protein coating significantly promoted HAEC migration.

[0036] Figure 3: CCN5 recombinant protein-coated stents demonstrate excellent safety. A: Scanning electron microscopy demonstrates good surface smoothness of the CCN5 recombinant protein-coated stent (CCN5rp). B: The CCN5 recombinant protein-coated stent and a simple PDA stent (control group) were placed in a purified, uniform red blood cell suspension and gently shaken at room temperature for 1 hour. The absorbance of the supernatant was measured spectrophotometrically. A 0.1% triton solution (which causes all red blood cells to lyse) was used as a control group to measure the hemolysis rate. The results showed that the CCN5 recombinant protein-coated stent and the simple PDA stent did not cause hemolysis. C: The CCN5 recombinant protein-coated stent was implanted into the right coronary artery of a pig during percutaneous coronary intervention (PCI). Coronary angiography performed preoperatively, immediately after the procedure, and seven days after the procedure demonstrated no significant intra-stent thrombosis caused by the CCN5 recombinant protein-coated stent.

[0037] Figure 4: CCN5 recombinant protein-coated stents promote endothelialization and inhibit in-stent restenosis. Bare metal stents (BMS) served as a control group, and sirolimus eluting stents (SES) served as a positive control group. Stents in each group were implanted into porcine coronary arteries. Optical coherence tomography (OCT) was performed 7 days after surgery to assess the extent of in-stent re-endothelialization. OCT and coronary angiography were performed 90 days after surgery to assess the extent of in-stent restenosis in vivo. Pig coronary arteries were sectioned and stained with hematoxylin and eosin (HE) for ex vivo observation and statistical analysis of the extent of in-stent restenosis. A: OCT results 7 days after PCI show that CCN5 recombinant protein-coated stents increase the intimal coverage of the stent struts and promote in-stent re-endothelialization. B: Coronary angiography 90 days after PCI shows that, similar to sirolimus eluting stents, CCN5 recombinant protein-coated stents inhibit in-stent stenosis. C: 90 days after PCI, optical coherence tomography (OCT) shows that, similar to sirolimus-eluting stents, the CCN5 recombinant protein-coated stent inhibits in-stent neointimal formation. D: 90 days after PCI, HE staining shows that, similar to sirolimus-eluting stents, the CCN5 recombinant protein-coated stent inhibits in-stent neointimal formation. Statistical graphs (from left to right) show neointimal area, in-stent restenosis rate, neovascular intima-media ratio, and vascular media area. DETAILED DESCRIPTION

[0038] After extensive and in-depth research, the inventors unexpectedly developed a medical stent comprising a support structure and a recombinant CCN5 protein. The surface of the support structure comprises a coating containing polydopamine, and the recombinant protein is covalently bound to the polydopamine, thereby being anchored to the support structure. The medical stent of the present invention is used to prevent in-stent restenosis after intravascular interventional therapy and to promote the proliferation and migration of human aortic endothelial cells (HAECs) and / or to promote endothelialization to inhibit in-stent restenosis. Based on this, the inventors completed the present invention.

[0039] the term

[0040] Polydopamine

[0041] Polydopamine is a biogenic amine compound with a polyphenol structure that has adhesiveness and biocompatibility, and is therefore widely used in the biomedical field.

[0042] CCN5 recombinant protein

[0043] CCN5 is an extracellular matrix protein, also known as WISP-2 (Wnt1 inducible signaling pathway protein-2), and a member of the extracellular matrix protein family. It is involved in various biological processes, including cell proliferation, cell migration, apoptosis, and cell differentiation. Recombinant CCN5 protein is a protein expressed and purified in large quantities through genetic engineering techniques and can be used to study the role of CCN5 in cell biology, physiology, and pathophysiology.

[0044] In a preferred embodiment, the sequence of the CCN5 recombinant protein of the present invention is shown as SEQ ID NO.1.

[0045] Endovascular interventional therapy

[0046] It is a method of treatment that is performed from inside the blood vessels through a catheter, including interventional surgery, stent implantation, balloon angioplasty, etc. This treatment method is commonly used to treat various vascular diseases such as coronary artery disease, cerebrovascular disease, and peripheral vascular disease.

[0047] Medical stents and their preparation

[0048] The "medical stent" described in the present invention is a stent that can promote the migration of human aortic endothelial cells (HAECs) and / or promote endothelialization to inhibit in-stent restenosis.

[0049] In the present invention, the medical stent comprises:

[0050] (a) a supporting structure, wherein a surface of the supporting structure comprises a coating, wherein the coating comprises polydopamine;

[0051] (b) CCN5 recombinant protein, wherein the recombinant protein is fixed to the support structure by binding to polydopamine.

[0052] In the present invention, the loading amount of the CCN5 recombinant protein per unit area of ​​the support structure determines the therapeutic effect of the medical stent. There is no particular limitation on the loading amount of the CCN5 recombinant protein per unit area of ​​the support structure, and a typical loading amount is 150-1000 ng / cm 2 , preferably, 250-800ng / cm 2 , more preferably, 300-500 ng / cm 2 A preferred loading capacity per unit area is when the loading capacity per unit area of ​​the CCN5 recombinant protein on the support structure is 300-500 ng / cm 2 When the stent is endothelialized, the stent is most effective in promoting endothelialization and inhibiting in-stent restenosis.

[0053] In the present invention, the medical stent is prepared by the following method:

[0054] (a) providing a support structure and CCN5 recombinant protein, wherein the surface of the support structure comprises a coating comprising polydopamine;

[0055] (b) incubating the support structure and the CCN5 recombinant protein under alkaline conditions, so that the quinone group of the polydopamine in the coating undergoes a Michael addition reaction with the amino group of the CCN5 recombinant protein under alkaline conditions, thereby coating the CCN5 recombinant protein on the support structure to obtain the medical stent.

[0056] The medical stent prepared by the present invention can promote the proliferation and migration of human aortic endothelial cells (HAECs) and / or promote endothelialization to inhibit in-stent restenosis, and can be used to prevent in-stent restenosis after intravascular interventional treatment.

[0057] The main advantages of the present invention include:

[0058] (1) The present invention is the first to develop a medical stent comprising a support structure and a CCN5 recombinant protein. The surface of the support structure comprises a coating comprising polydopamine, and the recombinant protein is covalently bound to the polydopamine and fixed to the support structure. The medical stent of the present invention is used to prevent in-stent restenosis after PCI, and is also used to promote the proliferation and migration of human aortic endothelial cells (HAECs), and / or promote endothelialization to inhibit in-stent restenosis.

[0059] (2) The present invention inhibits smooth muscle proliferation while promoting endothelial repair, thereby reducing long-term in-stent restenosis after PCI surgery.

[0060] (3) The present invention provides a CCN5 recombinant protein coated stent constructed by using a polydopamine coating as an intermediary to fix human CCN5 recombinant protein to a coronary artery metal stent through covalent bonding, which is used to prevent in-stent restenosis after PCI surgery.

[0061] (4) Human CCN5 recombinant protein was indirectly fixed to the surface of the metal coronary stent through a polydopamine coating to construct a CCN5 recombinant protein coated stent.

[0062] (5) The protein-coated stent is used to reduce the occurrence of in-stent restenosis after percutaneous coronary intervention (PCI).

[0063] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0064] Unless otherwise specified, the reagents and materials used in the examples of the present invention are commercially available products.

[0065] Example 1 CCN5rp coated stent manufacturing method and process

[0066] The CoCr disc and scaffold were immersed in a solution containing 2 mg / mL dopamine (10 mM Tris buffer, pH 8.5) at 25°C for 12 hours, removed, sonicated in PBS for 15 minutes, and then washed with water to remove weakly bound dopamine. Four consecutive dip coating steps were performed to form a four-layer PDA film, which was dried at 60°C for 1 hour.

[0067] The PDA membrane-coated CoCr disc and the scaffold were sterilized by ultraviolet irradiation for 1 hour and immersed in a solution containing human CCN5rp (SEQ ID NO. 1, purchased from PEPROTECH, product number 120-16) (1 μg / ml, 5% trehalose PBS buffer, pH = 8.0) at 37°C for 4 hours under sterile conditions to chemically fix CCN5rp on the PDA membrane.

[0068] Example 2 Detection of the loading capacity of CCN5rp coated stents

[0069] The chemical composition of the CCN5 coating was determined by X-ray photoelectron spectroscopy (XPS, Thermo Scientific K-Alpha, USA) (Figure 1A), and the loading density of CCN5rp on the CCN5-coated scaffold was verified in real time by a quartz crystal microbalance (QCM-D, Q-sense AB, Sweden) (Figure 1B).

[0070] As can be seen from the C1s high-resolution image in Figure 1A, after CCN5 was fixed on PDA, the peak intensities at 287.5eV and 284.1eV were weakened compared to the PDA sample, indicating that the quinone group in PDA was consumed during the reaction; the peak intensity at 285.6eV was weakened and moved to a high binding energy position, indicating that the aliphatic (aromatic) CN in PDA was consumed during the reaction; the peak area at 284.4eV increased, indicating that CCN5 containing a higher content of CH and CC was successfully attached. As can be seen from the N1s high-resolution spectrum in Figure 1A, the aliphatic CN / C=N peak area of ​​PDA-CCN5 is reduced relative to the PDA sample, the aromatic CN peak area is increased, and the binding energy is shifted to the binding energy, which may be caused by the abundant peptide bonds with CCN-5; the C-NH3 at 400.10eV + After CCN-5, the shift to 401.68 eV can be attributed to the transfer of protons from acids (catechol, C-OH, COOH) to amines (C-NH2). The above results indicate that the CCN5 recombinant protein was successfully loaded onto the surface of the metal scaffold or circular metal sheet (A). After rinsing with PBS, the metal crystal mass no longer changed and the loading capacity corresponding to the plateau was 330 ng / cm 2 Left and right (indicated by the red arrow), showing that the loading capacity of CCN5 recombinant protein is about 330ng / cm 2 (B).

[0071] Example 3 In vitro evaluation of CCN5 recombinant protein-coated CoCr discs and scaffolds

[0072] Human aortic endothelial cells (HAECs) PCS-100-011 TM ) with 1×10 5 Cells were seeded at a density of 10 cells / well onto CCN5rp-coated CoCr discs and cultured in endothelial cell culture medium (Sciencell, Cat No. 1001, USA) for 4 hours. Non-adherent cells were washed with PBS to remove them. Adherent cells were stained with phalloidin (1:200, Cat No. 40735ES75, Yeasen, China) and Hoechst (10 μg / ml). Cell number was counted in five random fields under a fluorescence microscope to observe cell adhesion to the membrane (Figure 2A).

[0073] Human aortic endothelial cells (HAECs) were cultured at 1×10 5 The cells were seeded at a density of 100 cells / well onto CCN5rp-coated CoCr plates, and the corresponding spectrophotometric values ​​were measured using the MTT assay at 0, 24, and 48 hours to calculate the cell proliferation capacity ( FIG2B ).

[0074] Human aortic endothelial cells (HAECs) were cultured at 1×10 5 The cells were seeded at a density of 100 cells / well on CCN5rp-coated CoCr plates and cultured in serum-free culture for 24 hours, followed by stimulation with 10% fetal bovine serum. After 6 hours, the cells were fixed with 4% paraformaldehyde and stained with Ki67 immunofluorescence to calculate the proportion of endothelial cells entering the proliferative phase ( Figure 2C ).

[0075] Human aortic endothelial cells (HAECs) stably overexpressing green fluorescent protein were cultured at a rate of 1×10 6 Cells were seeded at a density of 100 cells / well on CCN5rp-coated CoCr discs. After starvation for 24 h, the scratch area was observed using a fluorescence microscope at 24 h by a scratch test, and the cell migration ability was calculated (Figure 2D).

[0076] The results in Figure 2 show that the circular metal sheet coated with CCN5 recombinant protein can promote the adhesion (A), proliferation (B, C) and migration (D) of human aortic endothelial cells (HAECs).

[0077] Example 4 Surface Smoothness and Biocompatibility Analysis of CCN5rp-Coated CoCr Discs and Stents

[0078] The surface morphology of the PDA-coated and CCN5-coated stents was observed using a scanning electron microscope (SEM, ZEISS GeminiSEM 300, Germany) ( FIG3A ).

[0079] A quantitative hemolysis test was further performed to analyze the biocompatibility of the coated stent. Briefly, whole blood was gently mixed with a pH 7.4 PBS solution at a ratio of 1:2 and centrifuged at 3000 rpm for 6 minutes at room temperature to obtain red blood cells. The above steps were repeated 5 times to completely remove the serum from the red blood cell suspension. The 10-fold diluted red blood cells were placed in a 24-well plate as a 5% hematocrit solution. PDA-coated CoCr discs and CCN5rp-coated CoCr discs were placed in the corresponding 24-well plate wells, incubated on a shaker (medium speed) at 37°C for 1 hour, and then centrifuged at 6000 rpm for 5 minutes. The supernatant was collected and the hemoglobin absorbance displayed when the excitation light was 540 nm was recorded on an ELISA plate instrument (Tecan Infinite M200). The hemolysis percentage (HP) of each sample was calculated using the following formula: HP (%) = At ​​- Anc / Apc - Anc, where At is the absorbance of the test sample, Apc is the absorbance of the positive control (0.1% Triton), and Anc is the absorbance of the negative control (PBS). (Figure 3B)

[0080] Finally, in the porcine coronary artery model, we performed percutaneous coronary stent implantation through the porcine femoral artery access port and observed blood flow and thrombosis in the corresponding vascular segment of the stent by angiography immediately after stent implantation and on day 7 after surgery (Figure 3C).

[0081] The results in Figure 3 show that the CCN5 recombinant protein-coated stent has good safety, as demonstrated by good surface flatness (A), no hemolysis (B), and no intra-stent thrombosis before, immediately after, and on the seventh day after PCI in pigs (C).

[0082] Example 5: Verification of the effectiveness of CCN5rp coated stents through in vivo experiments in porcine coronary arteries

[0083] Five Chinese domestic pigs weighing 30 to 35 kg were provided by Shanghai Jiagan Biotechnology Co., Ltd. (Shanghai, China) and fed a standard laboratory diet (purchased from Shanghai Jiagan Biotechnology Co., Ltd.) for 7 days before the experiment. Dual antiplatelet therapy consisted of 100 mg of aspirin and 75 mg of clopidogrel daily for 3 days before surgery and throughout the study. Animal husbandry and all in vivo experiments were performed at Shanghai Anzhu Medical Technology Co., Ltd. (Shanghai, China).

[0084] On the day of the experiment, all animals were preanesthetized with an intramuscular injection of tiletamine, zolazepam (25 mg / kg), and xylazine (2 mg / kg). Mechanical ventilation was established, and anesthesia was maintained with inhaled isoflurane (1-2%) during the procedure. After local disinfection, the femoral artery was punctured, and a No. 6 arterial sheath (Terumo, Japan) was inserted. An intravenous bolus of 5000 UI of unfractionated heparin was administered. Digital subtraction angiography (OptimaCL323i, GE, USA) was used to locate the three major epicardial coronary arteries.

[0085] Each animal was randomly assigned to receive a CCN5rp-coated stent, a sirolimus-coated stent, or a bare-metal CoCr stent implanted in the left anterior descending artery (LAD), left circumflex artery (LCX), and right coronary artery (RCA). Quantitative coronary angiography (QCA) was used to measure coronary artery diameters after angiography. Stents were placed in the main coronary artery territory, with a stent-to-vessel ratio of 1.1 to 1.2:18. After cannulation, hemostasis was achieved by manual pressure, and the sheath was removed. The animals were allowed to recover from anesthesia.

[0086] Optical coherence tomography (OCT) was performed 7 days after surgery to assess the intimal coverage of the stent (Figure 4A). Coronary angiography and OCT were performed 90 days after surgery to observe and calculate the extent of in-stent neointimal thickening (Figures 4B-C). At the end of the final procedure, the animals were euthanized under deep anesthesia with a lethal dose of potassium chloride. The hearts were harvested, and the coronary arteries, at least 10 mm proximal and distal to the stent, were carefully dissected. After fixation, tissue sections were prepared and stained with hematoxylin and eosin (HE) to assess the severity of in-stent neointimal hyperplasia (Figure 4D).

[0087] The results in Figure 4 show that CCN5 recombinant protein coated stents promote epithelialization within 7 days after PCI in pigs (A) and inhibit in-stent restenosis 90 days after PCI in pigs (BD).

[0088] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A medical stent, characterized in that, The stent comprises: (a) A support structure, the surface of the support structure contains a coating, and the coating contains polydopamine; (b) CCN5 recombinant protein, which is fixed to the support structure by binding to polydopamine.

2. The medical stent according to claim 1, characterized in that, The recombinant protein covalently binds to polydopamine and is fixed to the support structure.

3. The medical stent according to claim 2, characterized in that, The support structure is selected from the group consisting of: coronary artery stent, carotid artery stent, abdominal aortic stent, femoral artery stent, renal artery stent, pulmonary artery stent, cerebral artery stent, or a combination thereof.

4. The medical stent according to claim 1, wherein The quinone group of polydopamine in the coating undergoes a Michael addition reaction with the amino group of CCN5 recombinant protein under alkaline conditions, thereby coating the CCN5 recombinant protein on the support structure.

5. The medical stent according to claim 1, characterized in that, The sequence of the CCN5 recombinant protein is as shown in SEQ ID NO.

1.

6. The medical stent according to claim 1, characterized in that, The loading amount of the CCN5 recombinant protein is 150 - 1000 ng / cm 2 , preferably 250 - 800 ng / cm 2 , more preferably 300 - 500 ng / cm 2 .

7. The medical stent according to claim 1, characterized in that, The number of layers of polydopamine coated on the support structure is 1 - 6 layers, preferably 2 - 5 layers; more preferably 3 - 4 layers.

8. The medical stent according to claim 1, characterized in that, The weight ratio of polydopamine to CCN5 recombinant protein in the medical stent is 0.1% - 2%, more preferably 0.2% - 1.5%, and most preferably 0.5% - 1%.

9. A manufacturing method of a medical stent, characterized in that, Comprising the steps: (a) Provide a support structure and CCN5 recombinant protein, the surface of the support structure contains a coating, and the coating contains polydopamine; (b) Incubate the support structure and CCN5 recombinant protein under alkaline conditions, such that the quinone group of polydopamine in the coating undergoes a Michael addition reaction with the amino group of CCN5 recombinant protein under alkaline conditions, thereby coating the CCN5 recombinant protein on the support structure to obtain the medical stent.

10. Use of the medical stent according to claim 1, characterized in that, For preparing a medical device for preventing in-stent restenosis after endovascular interventional therapy.

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