Transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as well as preparation method and application thereof

Abstract

The invention discloses a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as well as a preparation method and application thereof. Transition metal tetraphenylporphyrin and cobalt ions are accurately loaded in a covalent organic framework, and the coordination mode of the cobalt ions is optimized into O-O coordination, so that the catalytic stability of the transition metal tetraphenylporphyrin in a polymer material can be ensured while continuous and stable release of the cobalt ions is realized; the problems of short catalytic half-life caused by explosive release of transition metal ions and heavy metal poisoning caused by free transition metal ions are effectively avoided, and dynamic regulation and control and self-adaptive processes of NO release are realized through spatial precise configuration of two metal components in a covalent organic framework; the NO release rate is always maintained in an ideal physiological range, and the biological safety and the function durability of the material are improved.

Description

Technical Field

[0001] This application relates to the field of metal-organic framework materials technology, and in particular to a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material, its preparation method and application. Background Technology

[0002] Covalent organic frameworks (COFs) are a class of porous crystalline organic materials composed of light elements (C, O, N, B, etc.) linked by covalent bonds. They possess unique porous structures, high specific surface areas, and excellent chemical and thermal stability. Nitric oxide (NO) is an autocrine and paracrine signaling molecule whose main physiological function is to promote vascular homeostasis. It can promote endothelial cell adhesion and migration, inhibit smooth muscle contraction and growth, prevent platelet aggregation, and prevent leukocyte-endothelial cell adhesion. Current blood contact material coatings often employ the introduction of transition metals or organometallic complexes to catalyze substrates in the blood, such as S-nitrosoglutathione (GSNO), to achieve exogenous NO release in the early stages of implantation, thereby playing a role in antithrombosis and promoting endothelialization. However, this method relies entirely on transition metal ions to catalyze NO release, and the catalytic sites can be masked and blocked by non-specific adsorption of proteins and blood cells, as well as the migration of adjacent cells, leading to unstable NO release rates and hindering the long-term patency of the blood contact material. Summary of the Invention

[0003] This application provides a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material, its preparation method, and its application, which can achieve complementary release of endogenous and exogenous NO.

[0004] In a first aspect, this application provides a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material, comprising repeating units as shown in formula (I).

[0005]

[0006] Wherein, X is selected from Cu 2+ Ni 2+ Fe 2+ Zn 2+ and Co 2+ One of them;

[0007] R1 and R2 are each independently selected from one of -H, alkyl, hydroxyl, halogen, aryl, cycloalkyl, hydroxyalkyl, alkoxy, thioalkoxy, nitro, cyano, and amino.

[0008] In any embodiment of this application, the specific surface area of ​​the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material is 510 m².2 ·g -1 -520m 2 ·g -1 ; and / or, the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material comprises multiple pores with an average pore size of 2.00 nm to 2.20 nm.

[0009] In a second aspect, this application provides a method for preparing a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect, comprising: mixing 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal, 2,3-dihydroxyterephthalaldehyde, a first solvent and a catalyst, and preparing a covalent organic framework material intermediate by Schiff base condensation reaction; mixing the covalent organic framework material intermediate with cobalt nitrate hexahydrate and a second solvent, and carrying out a coordination reaction to obtain the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material.

[0010] In any embodiment of this application, the first solvent includes at least one of o-dichlorobenzene, m-dichlorobenzene, p-dichlorobenzene, toluene, xylene, n-butanol, methanol, ethanol, benzyl alcohol, phenethyl alcohol, and phenylpropanol; and / or, the catalyst includes at least one of acetic acid, formic acid, hydrochloric acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, and p-toluenesulfonic acid; and / or, the second solvent includes at least one of n-butanol, methanol, ethanol, and water.

[0011] In any embodiment of this application, the molar ratio of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal to 2,3-dihydroxyterephthalaldehyde is (0.4-0.6):1; and / or, the molar concentration of the catalyst is 5-7 mol / L; and / or, the mass ratio of the covalent organic framework material intermediate to cobalt nitrate hexahydrate is (0.04-0.06):1; and / or, the reaction temperature of the Schiff base condensation reaction is 110℃-130℃; and / or, the reaction time of the Schiff base condensation reaction is 60h-80h; and / or, the reaction temperature of the coordination reaction is 90℃-100℃; and / or, the reaction time of the coordination reaction is 20h-30h.

[0012] Thirdly, this application provides a medical material comprising a functional component and a matrix material, wherein the functional component comprises a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect or a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method described in the second aspect.

[0013] In any embodiment of this application, the medical material includes implantable materials or in vitro contact materials; preferably, implantable materials include vascular implants and non-vascular implants; preferably, vascular implants include at least one of artificial blood vessels and vascular stents; preferably, non-vascular implants include at least one of heart valves, tissue-engineered scaffolds, catheters and implantable leads; preferably, in vitro contact materials include dialysis tubing.

[0014] In any embodiment of this application, the medical material further includes a substrate material; and / or, the substrate material includes at least one of polydopamine, gelatin, chitosan, hyaluronic acid, and polyethyleneimine; and / or, the substrate material includes at least one of polymeric materials, metals and alloys, ceramics and composites, medical glass, carbon-based materials, and bio-based materials.

[0015] Fourthly, this application provides the application of a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect or a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method described in the second aspect in the preparation of biomedical materials for achieving complementary release of endogenous NO and exogenous NO. The biomedical materials include one or more of the following: (1) medical coating materials, including the aforementioned medical materials; (2) medical devices, including medical coating materials; (3) drug carriers; (4) nanocatalysts; preferably, the medical devices include implantable materials or in vitro contact materials.

[0016] In any embodiment of this application, the method for preparing a medical device includes: dissolving a precursor material and a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material in a buffer solution to obtain a precursor solution; coating the precursor solution onto the surface of a substrate material to obtain a medical device coated with a medical coating material; wherein the precursor material includes at least one of dopamine, gelatin, chitosan, hyaluronic acid, and polyethyleneimine, and the coating method includes at least one of spraying, spin coating, dip coating, physical adsorption, chemical grafting, cross-linking curing, sol-gel method, blend coating method, and layer-by-layer self-assembly method.

[0017] The transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material in this application embodiment successfully constructs a stable bimetallic covalent organic framework polymer material by precisely loading transition metal tetraphenylporphyrin and cobalt ions into COFs. In this polymer material, transition metal tetraphenylporphyrin is embedded in the COF structure in a firmly coordinated manner, continuously and efficiently catalyzing the decomposition of substrates such as GSNO in the blood to release NO, effectively avoiding the problem of short catalytic half-life caused by the explosive release of transition metal ions. At the same time, the cobalt ions bound in an OO coordination manner ensure the continuous and stable release of cobalt ions in vivo, which helps to activate the transcription factor HIF-1α signaling pathway and further promote the production of endogenous NO in endothelial cells, effectively avoiding the problem of decreased catalytic function of single transition metal ions due to endothelial cell migration and adhesion of components such as proteins in the blood. Through the precise spatial configuration of the two metal components in the COFs, the dynamic regulation and adaptive process of NO release is realized, ensuring that the NO release rate is always maintained within the ideal physiological range, thus improving the biosafety and functional durability of the material. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments of this application will be briefly introduced below. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 The results are XRD test results of 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1 of this application.

[0020] Figure 2 The results are BET test results of 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1 of this application.

[0021] Figure 3 The results are FTIR test results of 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1 of this application.

[0022] Figure 4 These are the XPS test results of 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1 of this application.

[0023] Figure 5 This is a schematic diagram of the vascular grafts of this application (wherein, from left to right, are actual images of PCL fiber tubes with different coatings prepared in Example 3, Example 4 and Example 2).

[0024] Figure 6This is a schematic diagram of the adhesion and proliferation of human umbilical vein endothelial cells (HUVECs) and human umbilical vein smooth muscle cells (HUVSMCs) on different coatings according to embodiments of this application (wherein, the left column is a schematic diagram of the adhesion and proliferation of HUVECs at different times; the right column is a schematic diagram of the adhesion and proliferation of HUVSMCs at different times).

[0025] Figure 7 This is a schematic diagram showing the NO release of different coatings in an embodiment of this application.

[0026] Figure 8 This is a schematic diagram of cell viability and mortality staining results in different coatings according to embodiments of this application.

[0027] Figure 9 This is a schematic diagram of the hemolysis test results corresponding to each group of coating samples in the embodiments of this application.

[0028] Figure 10 This is a schematic diagram showing the cobalt ion release of each group of coating samples in the embodiments of this application.

[0029] Figure 11 The various groups of coating samples in the embodiments of this application are subjected to Co loading. 2+ Schematic diagram of the change in specific surface area before and after (Figure A is a schematic diagram of the specific surface area test results of 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1; Figure B is a schematic diagram of the specific surface area test results of 2,5-DHTA COF and Co-2,5-DHTA COF prepared in Comparative Example 1; Figure C is a schematic diagram of the surface area test results of Bpy COF and Co-Bpy COF prepared in Comparative Example 2). Detailed Implementation

[0030] To make the purpose, technical solution, and beneficial technical effects of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the implementation details described in this specification are merely for illustrative purposes and are not intended to limit the scope of this application.

[0031] For simplicity, this application only explicitly discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form a range not explicitly stated; and any lower limit can be combined with other lower limits to form a range not explicitly stated, just as any upper limit can be combined with any other upper limit to form a range not explicitly stated. Furthermore, although not explicitly stated, every point or individual value between the endpoints of the range is included within that range. Therefore, each point or individual value can be used as its own lower or upper limit and combined with any other point or individual value or with other lower or upper limits to form a range not explicitly stated.

[0032] It should be noted that the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0033] Unless otherwise stated, the values ​​of the parameters mentioned in this application can be measured using various measurement methods commonly used in the art (e.g., they can be tested according to the methods given in the embodiments of this application). Unless otherwise stated, the test temperature for all parameters mentioned in this application is 25°C and the test pressure is standard atmospheric pressure.

[0034] The foregoing description of this application is not intended to describe every disclosed implementation or method. Instead, the following description provides more specific examples of exemplary embodiments. Throughout the application, guidance is provided through a series of embodiments, which can be used in various combinations. The examples listed are representative only and should not be construed as exhaustive.

[0035] The term "C1-C10 alkyl" refers to saturated straight-chain or branched alkanes with 1-12 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, C4 saturated straight-chain alkanes, C5 saturated straight-chain alkanes, C6 saturated straight-chain alkanes, C7 saturated straight-chain alkanes, C8 saturated straight-chain alkanes, C9 saturated straight-chain alkanes, and C1-C10 alkyl alkanes. 10 Saturated straight-chain alkanes, C4 saturated branched-chain alkanes, C5 saturated branched-chain alkanes, C6 saturated branched-chain alkanes, C7 saturated branched-chain alkanes, C8 saturated branched-chain alkanes, C9 saturated branched-chain alkanes, C 10 Saturated branched alkanes.

[0036] Existing blood contact material coatings often employ the introduction of transition metals or organometallic complexes to catalyze the formation of NO from substrates such as GSNO in the blood. These materials are prone to rapid degradation in vivo, causing explosive releases of metal ions and potentially leading to poisoning. Furthermore, the catalytic sites on the coating surface are inevitably masked and blocked by non-specific adsorption of proteins and blood cells, as well as the migration of adjacent vascular endothelial cells after implantation, resulting in a significant reduction in NO release capacity. Although endothelial cells possess endogenous NO secretion capacity, newly attached or migrating endothelial cells on the implant surface initially have poor function and cannot compensate for the insufficient NO supply caused by catalytic site shielding. Traditional coatings neglect the need to promote rapid recovery of endothelial cell function, making it difficult to achieve sustained, stable NO release and long-term patency. In view of the above-mentioned technical problems, the inventors conducted extensive experiments and developed a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material. As a surface coating component of blood contact materials, it can significantly reduce the risk of explosive release of metal ions and toxicity. It effectively overcomes the problems of insufficient NO release after the coating catalytic sites are covered by endothelial cells and slow recovery of function of newly formed endothelial cells. It achieves the complementary effect and dynamic regulation of endogenous and exogenous NO release, which is beneficial to the long-term patency of blood contact materials.

[0037] A first aspect of this application provides a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material, comprising repeating units as shown in formula (I).

[0038]

[0039] Wherein, X is selected from Cu 2+ Ni 2+ Fe 2+ Zn 2+ and Co 2+ One of them;

[0040] R1 and R2 are each independently selected from one of -H, alkyl, hydroxyl, halogen, aryl, cycloalkyl, hydroxyalkyl, alkoxy, thioalkoxy, nitro, cyano, and amino.

[0041] Preferably, X is Cu 2+ .

[0042] As an example, the alkyl group can be a C1-C10 alkyl group.

[0043] As an example, halogens can be -F, -Cl, -Br, or -I.

[0044] Preferably, both R1 and R2 are -H.

[0045] By precisely loading transition metal tetraphenylporphyrin and cobalt ions into COFs and optimizing the coordination mode of cobalt ions to OO coordination, it is possible to ensure the catalytic stability of transition metal tetraphenylporphyrin in polymer materials while achieving continuous and stable release of cobalt ions, effectively avoiding the catalytic instability of transition metal ions (such as Cu). 2+ Ni 2+ Fe 2+ Zn 2+ and Co 2+ The explosive release of NO leads to a short catalytic half-life and the problem of heavy metal poisoning caused by free transition metal ions. The transition metal tetraphenylporphyrin (TPP) in the cobalt covalent organic framework polymer material catalyzes the release of NO from substrates such as GSNO in the blood, mimicking endothelial cell function to inhibit platelet activation, smooth muscle cell proliferation, and inflammatory responses, thus promoting endothelialization. Simultaneously, cobalt ions, by mimicking a hypoxic state, activate the HIF-1α signaling pathway, thereby stimulating the secretion of endothelial pro-angiogenic factor (VEGF) and the expression of endothelial nitric oxide synthase (enos), promoting endogenous NO release. This effectively avoids the problem of decreased catalytic function of a single transition metal ion due to endothelial cell migration and adhesion of proteins and other components in the blood. Through precise spatial configuration of the two metal components in the COFs, dynamic regulation and adaptive processes of endogenous and exogenous NO release are achieved, ensuring that NO release remains within an effective range.

[0046] In some embodiments, the specific surface area of ​​the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material is 510 m². 2 ·g -1 -520m 2 ·g -1 ; and / or, the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material comprises multiple pores with an average pore size of 2.00 nm to 2.20 nm.

[0047] As an example, the specific surface area of ​​a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material can be 510 m². 2 ·g -1 511m 2 ·g -1 512m 2 ·g -1 513m 2 ·g -1 514m 2 ·g -1 515m 2 ·g -1 516m 2 ·g -1 517m 2 ·g -1518m 2 ·g -1 519m 2 ·g -1 Or 520m 2 ·g -1 .

[0048] Optionally, the specific surface area of ​​the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material is 517 m². 2 ·g -1 .

[0049] As an example, the average pore size of the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material can be 2.00 nm, 2.01 nm, 2.02 nm, 2.03 nm, 2.04 nm, 2.05 nm, 2.06 nm, 2.07 nm, 2.08 nm, 2.09 nm, 2.10 nm, 2.11 nm, 2.12 nm, 2.14 nm, 2.16 nm, 2.18 nm, or 2.20 nm.

[0050] Optionally, the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material has an average pore size of 2.04 nm.

[0051] The second aspect of this application provides a method for preparing a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect, comprising: mixing 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal, 2,3-dihydroxyterephthalaldehyde, a first solvent and a catalyst, and preparing a covalent organic framework material intermediate by Schiff base condensation reaction; mixing the covalent organic framework material intermediate with cobalt nitrate hexahydrate and a second solvent, and carrying out a coordination reaction to obtain the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material.

[0052] As an example, the transition metal could be copper, iron, nickel, cobalt, or zinc.

[0053] The transition metal in the tetraphenylporphyrin ring has a valence state of +2.

[0054] As an example, 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal can be 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-copper, 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-nickel, 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-iron, 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-cobalt, or 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-zinc. In some embodiments, the first solvent includes at least one of o-dichlorobenzene, m-dichlorobenzene, p-dichlorobenzene, toluene, xylene, n-butanol, methanol, ethanol, benzyl alcohol, phenethyl alcohol, and phenylpropanol; and / or, the catalyst includes at least one of acetic acid, formic acid, hydrochloric acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, and p-toluenesulfonic acid; and / or, the second solvent includes at least one of n-butanol, methanol, ethanol, and water.

[0055] Optionally, the first solvent includes o-dichlorobenzene and n-butanol.

[0056] Optionally, the volume ratio of o-dichlorobenzene to n-butanol is 1:1.

[0057] Optionally, the catalyst includes acetic acid.

[0058] The second solvent includes ethanol and water.

[0059] Optionally, the volume ratio of ethanol to water is 1:1.

[0060] In some embodiments, the molar ratio of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal to 2,3-dihydroxyterephthalaldehyde is (0.4-0.6):1; and / or, the molar concentration of the catalyst is 5 mol / L-7 mol / L; and / or, the mass ratio of the covalent organic framework material intermediate to cobalt nitrate hexahydrate is (0.04-0.06):1; and / or, the reaction temperature of the Schiff base condensation reaction is 110℃-130℃; and / or, the reaction time of the Schiff base condensation reaction is 60h-80h; and / or, the reaction temperature of the coordination reaction is 90℃-100℃; and / or, the reaction time of the coordination reaction is 20h-30h.

[0061] As an example, the molar ratio of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal to 2,3-dihydroxyterephthalaldehyde can be 0.4:1, 0.42:1, 0.45:1, 0.48:1, 0.5:1, 0.52:1, 0.54:1, 0.56:1, 0.58:1, or 0.6:1.

[0062] Optionally, the molar ratio of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal to 2,3-dihydroxyterephthalaldehyde is 0.5:1.

[0063] As an example, the molar concentration of the catalyst can be 5 mol / L, 5.5 mol / L, 6 mol / L, 6.5 mol / L, or 7 mol / L.

[0064] Optionally, the molar concentration of the catalyst can be 6 mol / L.

[0065] As an example, the mass ratio of the covalent organic framework material intermediate to cobalt nitrate hexahydrate can be 0.04:1, 0.045:1, 0.05:1, 0.055:1, or 0.06:1.

[0066] Optionally, the mass ratio of the covalent organic framework material intermediate to cobalt nitrate hexahydrate is 0.05:1.

[0067] As an example, the reaction temperature for Schiff base condensation can be 110°C, 115°C, 120°C, 125°C, or 130°C.

[0068] Optionally, the Schiff base condensation reaction is carried out at a temperature of 120°C.

[0069] As an example, the reaction time for Schiff base condensation reaction can be 60h, 65h, 68h, 70h, 71h, 72h, 73h, 74h, 75h, 78h or 80h.

[0070] Optionally, the Schiff base condensation reaction can be carried out over a period of 72 hours.

[0071] As an example, the reaction temperature for coordination reactions can be 90℃, 92℃, 93℃, 94℃, 95℃, 96℃, 98℃, or 100℃.

[0072] Optionally, the reaction temperature for the coordination reaction is 95°C.

[0073] As an example, the reaction time for coordination reactions can be 20h, 22h, 24h, 26h, 28h, or 30h.

[0074] Optionally, the reaction time for the coordination reaction is 24 hours.

[0075] Optionally, the Schiff base condensation reaction specifically includes: adding 0.06 mmol of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal and 0.12 mmol of 2,3-dihydroxyterephthalaldehyde (0.12 mmol) to a 10 mL Pyrex tube in succession with o-dichlorobenzene and n-butanol solution (1.5 mL / 1.5 mL). The mixture is then sonicated for 15 minutes to obtain a homogeneous dispersion. Next, 0.3 mL of 6 M acetic acid aqueous solution is added to the mixture, and the tube is rapidly frozen in a liquid nitrogen bath at 77 K, followed by three cycles of "freeze-evacuate-thaw". After degassing, the Pyrex tube is flame-sealed and reacted at 120 °C for 72 hours. After the reaction, the precipitate is collected by centrifugation, washed with anhydrous tetrahydrofuran and acetone, and then the product is vacuum-dried overnight at 60 °C to obtain a covalent organic framework intermediate.

[0076] Optionally, the coordination reaction specifically includes: adding a mixture of a covalent organic framework material intermediate (50.00 mg) and cobalt nitrate hexahydrate (1.00 g, 3.44 mmol) to an ethanol / water mixture (v / v = 1:1) and stirring at 95 °C for 24 hours. After the reaction is complete, the mixture is washed several times with water to ensure the removal of residual metal ions. Then, it is dried overnight in a vacuum oven at 60 °C to obtain a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material.

[0077] A third aspect of the embodiments of this application provides a medical material, including a functional component and a matrix material. The functional component includes a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect or a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method described in the second aspect.

[0078] Optionally, the aforementioned medical materials can be used as a base material, or as a functional coating, covering layer, or composite layer on various substrate materials.

[0079] In some embodiments, the medical material includes implantable materials or in vitro contact materials; preferably, implantable materials include vascular implants and non-vascular implants; preferably, vascular implants include at least one of artificial blood vessels and vascular stents; preferably, non-vascular implants include at least one of heart valves, tissue-engineered scaffolds, catheters and implantable leads; preferably, in vitro contact materials include dialysis tubing.

[0080] Optionally, medical materials include implantable materials.

[0081] Optionally, implantable materials include vascular implants.

[0082] Optionally, the vascular implant includes at least one of a small-diameter artificial blood vessel or a vascular stent.

[0083] In some embodiments, the medical material further includes a substrate material; and / or, the substrate material includes at least one of polydopamine, gelatin, chitosan, hyaluronic acid, and polyethyleneimine; and / or, the substrate material includes at least one of polymeric materials, metals and alloys, ceramics and composites, medical glass, carbon-based materials, and bio-based materials.

[0084] Among them, polymer materials include, but are not limited to, biodegradable plastics such as polysulfone (PSU), polyethersulfone (PES), polyethylene terephthalate, polyvinylidene fluoride (PVDF), polyurethane (PU), polyethylene (PE), polypropylene (PP), polytetrafluoroethylene, polyamide (PA), nylon, silicone rubber, polystyrene (PS), polylactic acid (PLA), and polycaprolactone (PCL); metal and alloy materials include, but are not limited to, stainless steel (such as 316L), cobalt-chromium alloys, platinum-chromium alloys, magnesium alloys, titanium and titanium alloys, and precious metals (such as gold and silver); ceramics and composite materials include, but are not limited to, organic-inorganic composite materials such as alumina, zirconium oxide, and hydroxyapatite; carbon-based materials include, but are not limited to, carbon fibers and graphene derivatives; and bio-based materials include, but are not limited to, silk fibroin and collagen.

[0085] Optionally, the matrix material includes polydopamine.

[0086] Optionally, the base material includes polycaprolactone.

[0087] Alternatively, the substrate material can be a vascular graft prepared by electrospinning, such as an artificial blood vessel or vascular stent, tissue engineering scaffold, nanofiber dressing, etc.

[0088] In some embodiments, the substrate material is an artificial blood vessel prepared using an electrospinning process.

[0089] The fourth aspect of this application provides the application of a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in the first aspect or a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method described in the second aspect in the preparation of biomedical materials for achieving complementary release of endogenous NO and exogenous NO. The biomedical materials include one or more of the following: (1) medical coating materials, including the aforementioned medical materials; (2) medical devices, including medical coating materials; (3) drug carriers; (4) nanocatalysts; preferably, the medical devices include implantable materials or in vitro contact materials.

[0090] As examples, medical coating materials can be thin-film coatings, nanoparticle coatings, and composite coatings such as composite antibacterial coatings or composite anti-corrosion coatings. Thin-film coatings involve fabricating transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials into nanofilms or ultrafilms, which are then coated onto a substrate surface using methods such as spin coating, spraying, dip coating, or chemical vapor deposition (CVD). Nanoparticle coatings involve preparing transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials into nanoparticles, dispersing them in a solvent to form a suspension, and then coating them onto a substrate using methods such as spraying, dip coating, or printing. Composite coatings involve combining transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials with other materials (such as polymers, metal oxides, carbon materials, etc.) to form a multifunctional coating.

[0091] As an example, implantable materials include vascular implants and non-vascular implants; vascular implants include at least one of artificial blood vessels and vascular stents; non-vascular implants include at least one of heart valves, tissue-engineered scaffolds, catheters and implantable leads.

[0092] As an example, in vitro contact materials include dialysis tubing.

[0093] As an example, a drug carrier can load at least one of the following pharmaceutical active ingredients:

[0094] (1) Antithrombotic drugs, including but not limited to: anticoagulants such as heparin and low molecular weight heparin; antiplatelet drugs such as pioglitazone and aspirin;

[0095] (2) Drugs that promote vascular or endothelial repair, including but not limited to: vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF);

[0096] (3) Anti-inflammatory, anti-fibrotic, or antioxidant drugs, including but not limited to: dexamethasone, triamcinolone (glucocorticoids, anti-inflammatory); N-acetylcysteine ​​(NAC), glutathione (antioxidants);

[0097] (4) Antibacterial or anti-infective drugs, including but not limited to: antibiotics such as vancomycin, cephalosporins, and gentamicin (antimicrobial peptides); and non-antibiotics such as nano-silver and quaternary ammonium salts;

[0098] (5) Other functional drugs, including but not limited to: NO precursors or NO donor compounds such as sodium nitroprusside (SNP) and GSNO, which can synergistically release NO with transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials; RNA, siRNA or CRISPR vectors, which are used for gene therapy and intervention.

[0099] As examples, nanocatalysts can be supported catalysts, composite catalysts, biocatalysts, etc. Among them, supported catalysts are composite catalysts formed by loading transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials onto a support (such as silica, carbon materials, or metal oxides); composite catalysts are multifunctional catalysts formed by combining transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials with other catalytic materials (such as metal nanoparticles or semiconductor materials); and biocatalysts are bio-inorganic composite catalysts formed by combining transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer materials with enzymes or biomolecules.

[0100] The aforementioned medical coating materials, medical devices, drug carriers, and nanocatalysts can synergistically release endogenous and exogenous nitric oxide, ensuring the persistence and stability of NO release.

[0101] In some embodiments, the preparation method of the medical device includes: dissolving a precursor material and a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material in a buffer solution to obtain a precursor solution; coating the precursor solution onto the surface of a substrate material to obtain a medical device with a medical coating material on its surface; wherein the precursor material includes at least one of dopamine, gelatin, chitosan, hyaluronic acid and polyethyleneimine, and the coating method includes at least one of spraying, spin coating, dip coating, physical adsorption, chemical grafting, crosslinking curing and sol-gel method, blend coating method, and layer-by-layer self-assembly method.

[0102] Alternatively, the coating method may include dip coating.

[0103] The bimetallic covalent organic framework material of the present invention combines the functionality of transition metals, especially copper and cobalt, and is applied to the coating of various vascular implants such as small-diameter artificial blood vessels and vascular stents. It achieves complementary and sequential effects of endogenous and exogenous NO release, significantly improving the short-term and long-term patency of vascular implants. Compared with the use of expensive and easily inactivated biomolecules such as growth factors, the bimetallic COFs material of the present invention has significant advantages such as low cost, high stability and long-lasting function, making it suitable for large-scale production and clinical application.

[0104] Example

[0105] The following embodiments describe the disclosure of this application in more detail. These embodiments are for illustrative purposes only, as various modifications and variations will be apparent to those skilled in the art within the scope of the disclosure of this application. Unless otherwise stated, all parts, percentages, and ratios reported in the following embodiments are based on mass, and all reagents used in the embodiments are commercially available or synthesized by conventional methods and can be used directly without further processing, and the instruments used in the embodiments are commercially available.

[0106] Example 1: Preparation of Co-2,3-DHTA COF

[0107] Example 1 of this application provides a method for preparing a copper tetraphenylporphyrin-cobalt covalent organic framework polymer material, comprising the following steps:

[0108] Preparation of S1,2,3-DHTA COF

[0109] 0.06 mmol of 5,10,15,20-tetra-(4-aminophenyl)porphyrin-copper(II) and 0.12 mmol of 2,3-dihydroxyterephthalaldehyde (2,3-DHTA) were successively added to a 10 mL Pyrex tube along with 1.5 mL of o-dichlorobenzene solution and 1.5 mL of n-butanol solution. The mixture was sonicated for 15 minutes to obtain a homogeneous dispersion. Then, 0.3 mL of 6 M acetic acid aqueous solution was added to the mixture, and the tube was rapidly frozen in a liquid nitrogen bath at 77 K, followed by three cycles of "freeze-evacuate-thaw". After degassing, the Pyrex tube was flame-sealed and reacted at 120 °C for 72 hours. After the reaction, the precipitate was collected by centrifugation, carefully washed with anhydrous tetrahydrofuran and acetone, and then the product was vacuum-dried overnight at 60 °C to obtain black 2,3-DHTA COF.

[0110] Preparation of S2,Co-2,3-DHTA COF

[0111] A mixture of 50.00 mg of 2,3-DHTA COF and 1.00 g of cobalt nitrate hexahydrate (3.44 mmol) was added to an ethanol / water mixture (v / v = 1:1), and stirred at 95 °C for 24 hours. After the reaction was complete, the mixture was washed several times with water to ensure the removal of residual metal ions. It was then dried overnight in a vacuum oven at 60 °C to obtain Co-2,3-DHTA COF.

[0112] The reaction route is shown below:

[0113]

[0114] Example 2: Construction of PCL fiber material with PDA-Co-2,3-DHTA COF composite coating

[0115] Electrospun fiber materials were prepared by electrospinning 10% (w / v) polycaprolactone (PCL). First, PCL particles were dissolved in hexafluoroisopropanol (HFIP) and stirred at room temperature for 6 hours. Next, the solution was drawn into a syringe and attached to an electrospinning apparatus. The electrospinning parameters were set as follows: the distance between the syringe tip and the receiver was 20 cm; the syringe needle was connected to a 10 kV positive high voltage, the receiver plate was connected to a -3 kV negative high voltage, and the injection speed was 0.2 mL / min. -1 The electrospinning time was 30 min. Fibers were deposited on a rotating stainless steel mandrel with a diameter of 2.5 mm to prepare an electrospun fiber tube at a rotation speed of 400 rpm. Fibers were also deposited on a flat plate collector to prepare an electrospun fiber membrane. Residual solvents were removed by vacuum drying for at least 48 hours. 2 mg / mL of dopamine was dissolved in a Tris buffer solution at pH 8.5, and then 2 mg / mL of the nano-Co-2,3-DHTA COF prepared in Example 1 was immediately ultrasonically dispersed into the solution to obtain a mixed solution. Finally, the electrospun fiber tube and the electrospun fiber membrane were immersed in the mixed solution and shaken at 37°C for 12 hours each time. After each immersion, ultrasonic cleaning was performed to remove unfixed COFs from the surface. This process was repeated three times to prepare PCL fiber tubes and PCL fiber membranes with a PDA-Co-2,3-DHTA COF composite coating.

[0116] Example 3: Construction of PCL fiber material with PDA coating on surface

[0117] The only difference between this and Example 2 is the preparation of the mixed solution: 2 mg / mL of dopamine is dissolved in a Tris buffer solution with pH=8.5 to obtain a mixed solution. Finally, the electrospun fiber tube and the electrospun fiber membrane are immersed in the mixed solution and shaken at 37 degrees Celsius for 12 hours to prepare PCL fiber tubes and PCL fiber membranes with PDA coatings on their surfaces.

[0118] Example 4: Construction of PCL fiber material with PDA-2,3-DHTA COF composite coating

[0119] The only difference between Example 4 and Example 2 is that the bimetallic COF material Co-2,3-DHTA COF is replaced with the monometallic COF material 2,3-DHTA COF.

[0120] The preparation method of the single-metal COF material 2,3-DHTA COF includes the following steps.

[0121] 0.06 mmol of 5,10,15,20-tetra-(4-aminophenyl)porphyrin-copper(II) and 0.12 mmol of 2,3-dihydroxyterephthalaldehyde (2,3-DHTA) were successively added to a 10 mL Pyrex tube along with 1.5 mL of o-dichlorobenzene solution and 1.5 mL of n-butanol solution. The mixture was sonicated for 15 minutes to obtain a homogeneous dispersion. Then, 0.3 mL of 6M acetic acid aqueous solution was added to the mixture, and the tube was rapidly frozen in a liquid nitrogen bath at 77 K, followed by three cycles of "freeze-evacuate-thaw". After degassing, the Pyrex tube was flame-sealed and reacted at 120 °C for 72 hours. After the reaction, the precipitate was collected by centrifugation, carefully washed with anhydrous tetrahydrofuran and acetone, and then the product was vacuum-dried overnight at 60 °C to obtain a black 2,3-DHTA COF powder, which is the single-metal COF material.

[0122] Example 5: Construction of a hemodialysis tubing with a PDA-Co-2,3-DHTA COF composite coating

[0123] 2 mg / mL of dopamine was dissolved in a Tris buffer solution at pH 8.5. Then, 2 mg / mL of nano-Co-2,3-DHTA COF prepared in Example 1 was immediately ultrasonically dispersed into the above solution to obtain a mixed solution. Finally, the PVC hemodialysis tubing was immersed in the above mixed solution and shaken at 37 degrees Celsius for 12 hours each time. At the end of each time, ultrasonic cleaning was performed to remove the unfixed COFs on the surface. This process was repeated 3 times to prepare dialysis tubing with a PDA-Co-2,3-DHTA coating.

[0124] Example 6: Construction of an artificial valve with a Co-2,3-DHTA COF-gelatin composite coating.

[0125] Weigh gelatin and add it to deionized water at a concentration of 80 mg / ml. Stir magnetically in a 37°C water bath for 1 hour to completely dissolve the gelatin and form a clear solution. Add Co-2,3-DHTA COF nanoparticles to the gelatin solution at room temperature while stirring to prevent agglomeration. Perform ultrasonic treatment for 5–10 minutes to further improve dispersion uniformity, achieving a final concentration of 2 mg / ml. Take the polyurethane artificial heart valve substrate and ultrasonically clean it sequentially with anhydrous ethanol, deionized water, and NaOH solution to remove surface impurities. Immerse the pretreated polyurethane valve in the COF-gelatin composite solution at 37°C for 5–30 minutes to ensure full coating. After removal, dry in a 37°C oven for 30 minutes. Repeat the coating and drying process four times to increase the coating thickness. The coated valve was placed in a 0.1% glutaraldehyde aqueous solution and soaked for 2 minutes. After cross-linking, it was rinsed multiple times with PBS or deionized water to remove residual cross-linking agent and prevent toxic residue. Finally, it was dried in a vacuum oven at 37°C for 24 hours.

[0126] Example 7: Construction of a vascular stent with a surface coated with PDA-Co-2,3-DHTA

[0127] 2 mg / mL of dopamine was dissolved in a Tris buffer solution at pH 8.5. Then, 2 mg / mL of nano-Co-2,3-DHTA COF prepared in Example 1 was immediately ultrasonically dispersed into the above solution to obtain a mixed solution. Finally, the SS 316L coronary stent was immersed in the above mixed solution and shaken at 37 degrees Celsius for 12 hours each time. At the end of each time, ultrasonic cleaning was performed to remove the unfixed COFs on the surface. The process was repeated three times to prepare vascular stents with PDA-Co-2,3-DHTA coating.

[0128] Comparative Example 1: Preparation of Co-2,5-DHTA COF

[0129] The only difference between Comparative Example 1 and Example 1 is that 2,3-DHTA in step S1 is replaced with 2,5-dihydroxyterephthalaldehyde (2,5-DHTA), and the final product Co-2,5-DHTA COF is prepared.

[0130] The reaction route is shown below:

[0131]

[0132] Preparation of Comparative Example 2: Co-Bpy COF

[0133] The only difference between Comparative Example 2 and Example 1 is that 2,3-DHTA in step S1 is replaced with 2,2'-bipyridine (Bpy), cobalt nitrate hexahydrate in step S2 is replaced with cobalt acetate, and ethanol in step S2 is replaced with methanol, so as to prepare the final product Co-Bpy COF.

[0134] The reaction route is shown below:

[0135]

[0136] Comparative Example 3: Construction of PCL fiber material with PDA-Co-2,5-DHTA COF composite coating

[0137] The only difference between Comparative Example 3 and Example 2 is that the nano-Co-2,3-DHTA COF was changed to Co-2,5-DHTA COF.

[0138] Comparative Example 4: Construction of PCL fiber material with PDA-Co-Bpy COF composite coating

[0139] The only difference between Comparative Example 4 and Example 2 is that the nano-Co-2,3-DHTA COF was changed to Co-Bpy COF.

[0140] Performance testing

[0141] 1. X-ray diffraction (XRD), surface area measurement (BET), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS) were performed on the 2,3-DHTA COF and Co-2,3-DHTA COF prepared in Example 1, respectively. The test results are shown in the figure. Figures 1-4 ,in, Figure 1 To obtain the XRD patterns of 2,3-DHTA COF before and after Co, Figure 2 To obtain the BET spectrum of 2,3-DHTACOF before and after Co, Figure 3 To obtain the FTIR spectra of 2,3-DHTA COF before and after Co, Figure 4 XPS spectra of 2,3-DHTA COF before and after Co preparation are shown. These results indicate that Co-2,3-DHTA COF was successfully synthesized.

[0142] 2. Photographs were taken of the PCL fiber tubes (i.e., vascular grafts) with different coatings prepared in Examples 3, 4, and 2. Figure 5 The image shown is a picture of the actual product. From left to right, the product is a PCL fiber tube with a PDA coating, a PCL fiber tube with a PDA-2,3-DHTA COF composite coating, and a PCL fiber tube with a PDA-Co-2,3-DHTA COF composite coating.

[0143] 3. Cell adhesion and proliferation tests were performed on the PCL fiber membranes with PDA-Co-2,3-DHTA COF composite coatings prepared in Example 2, the PCL fiber membranes with PDA coatings prepared in Example 3, and the PCL fiber membranes with PDA-2,3-DHTA COF composite coatings prepared in Example 4. The test methods are as follows: HUVECs and HUVSMCs were divided into two groups: one group of cell culture medium lacking NO donor; the other group of cell culture medium containing 10 μM NO donor (GSNO) and 10 μM glutathione (GSH). The culture medium containing NO donor was changed daily. The PCL fiber membranes with different coatings were placed in confocal dishes, and 1 mL of 1×10⁻⁶ g / mL of glutathione (GSH) was added. 5 Cell culture medium for HUVECs or HUVSMCs was prepared and statically incubated in a cell culture incubator containing 5% CO2 at 37°C for 4 h, 1 d, and 3 d. After removal, the samples were washed with PBS (pH 7.4) and fixed with 2.5% glutaraldehyde. After calcein AM staining, the growth of HUVECs or HUVSMCs in PCL fiber membranes of each group was observed under a confocal microscope.

[0144] Depend on Figure 6 It is understood that the PCL fiber membrane of the present invention with a PDA-Co-2,3-DHTA COF composite coating (i.e. Figure 6 The CuCo group) and the PCL fiber membrane with a PDA-2,3-DHTA COF composite coating on its surface (i.e. Figure 6 In the presence of GSNO donors, the Cu group significantly promoted the adhesion and proliferation of HUVECs and inhibited the adhesion and proliferation of HUVSMCs. Among them, the PCL fiber membrane with a PDA-Co-2,3-DHTA COF composite coating (i.e., Figure 6 The CuCo group in this invention shows the best performance. The PCL fiber membrane of this invention, with a PDA-Co-2,3-DHTA COF composite coating (i.e., Figure 6 The CuCo group in COFs catalyzes the exogenous release of NO from substrates in the blood, promoting endothelialization and inhibiting HUVSMC proliferation. With endothelialization, cobalt ions can further promote enos expression and VEGF secretion through the HIF-1α signaling pathway, further promoting the functional recovery of HUVECs.

[0145] 4. NO release was detected in the PCL fiber membranes prepared in Example 2 and Example 4, both coated with a PDA-Co-2,3-DHTA COF composite coating. The test method was as follows: the Griess method was used to determine the stable metabolites of NO (nitrite, NO2). - The NO release capacity was assessed by measuring the concentration of [NO2] in aqueous solution. NO released by HUVECs and copper porphyrins is readily oxidized to nitrite (NO2) in aqueous solution. - ) and nitrates (NO3) - The total nitrite concentration in the cell culture medium was determined by the Griess method, which can indirectly reflect the changes in the total amount of NO released by HUVECs and the total amount of NO released by membrane catalysis.

[0146] Place the sample into a 24-well cell culture plate and add 1 mL of a solution containing 1×10⁻⁶ cells / well. 5 The cell culture medium was statically incubated in a cell culture incubator containing 5% CO2 at 37°C. Figure 6 The diagram illustrating the adhesion and proliferation of HUVECs and HUVSMCs on different coatings was used to determine the coverage area of ​​HUVECs and HUVSMCs on the coatings. At approximately 10%, 30%, 50%, and 70% cell coverage, the original culture medium was replaced with 1 ml of fresh cell culture medium containing 10 μM GSNO and 10 μM GSH. After incubation for 4 hours, the cell culture supernatant was collected. The amount of NO was quantified using a Griess reaction assay kit (Beyotime Biotechnology Research Institute, China). Absorbance was measured at 540 nm using a microplate reader. Changes in absorbance at 540 nm were recorded and used to calculate nitrite concentration, thereby calculating the NO release rate. The NO release rate was calculated using the following formula:

[0147]

[0148] OD1 and OD2 represent the absorbance at 540 nm at different times, K is the coefficient of absorbance at 540 nm related to nitrite concentration, V is the volume of the test solution, S is the area of ​​the vascular graft, and T1 and T2 represent different detection times.

[0149] Test results are available Figure 7 ,Depend on Figure 7 It can be seen that as the cell coverage area of ​​HUVECs increases, the PCL fiber membrane with a PDA-Co-2,3-DHTA COF composite coating (corresponding to) of the present invention... Figure 7The PCL-PDA-CuCo group can maintain the NO release within the effective range; while the PCL fiber membrane with a PDA-2,3-DHTA COF composite coating (corresponding to...) Figure 7 In the PCL-PDA-Cu group, as the coverage area of ​​HUVECs cells increased, the release rate of NO decreased significantly. Long-term use in blood contact materials can easily lead to intimal hyperplasia and calcification.

[0150] 5. The PCL fiber membranes with PDA-Co-2,3-DHTA COF composite coatings prepared in Example 2 and the PCL fiber membranes with PDA coatings prepared in Example 3 were subjected to cytotoxicity and blood compatibility tests. The test methods are as follows:

[0151] (1) Cytotoxicity:

[0152] The PCL fiber membranes with PDA-Co-2,3-DHTA COF composite coatings prepared in Example 2 and the PCL fiber membranes with PDA coatings prepared in Example 3 were placed in confocal dishes, and 1 ml of DMEM cell culture medium (containing 1×10⁻⁶ cells / mL) was added. 5 After incubating PCL fiber membranes with HUVECs for 1 day, the PCL fiber membranes were removed, washed with PBS, and then immersed in 1 mL of PBS (containing calcein (1:500) and propidium iodide (1:300)). After incubation at 37°C in the dark for 15 min, the membranes were fixed with paraformaldehyde (4.0%, w / v). Fluorescence images of HUVECs on the PCL fiber membranes were captured by laser scanning confocal microscopy.

[0153] Figure 8 The diagram shows the results of cell viability and mortality staining, regardless of whether the PCL fiber membrane is coated with a PDA-Co-2,3-DHTA COF composite coating (corresponding to...). Figure 8 PCL-PDA-COF group or PCL fiber membrane with PDA coating on surface (corresponding to Figure 8 After co-culturing with cells, the HuvECs and HuvSMCs adhering to the PCL-PDA group showed green fluorescence but no red fluorescence, indicating that the PCL fiber membrane with the PDA-Co-2,3-DHTA COF composite coating of the present invention is non-cytotoxic and has excellent cell compatibility.

[0154] (2) Blood compatibility

[0155] S1, Blood collection

[0156] Fresh anticoagulated whole blood was collected from healthy rabbits using EDTA anticoagulation tubes to separate red blood cells: The whole blood was centrifuged at 1000g for 10 minutes to remove the plasma and leukocyte layers. Red blood cells were washed: The red blood cells were washed three times with physiological saline or PBS buffer, centrifuged at 1000g for 5 minutes each time, until the supernatant was clear.

[0157] S2, Preparation of red blood cell suspension

[0158] The washed red blood cells are diluted with physiological saline or PBS to a certain concentration (usually 2% or 5% red blood cell suspension, v / v).

[0159] S3, Blood Compatibility Test

[0160] The PCL fiber membranes with PDA-Co-2,3-DHTA COF composite coatings prepared in Example 2, the PCL fiber membranes with PDA coatings prepared in Example 3, and the PCL fiber membranes with PDA-2,3-DHTA COF composite coatings prepared in Example 4 were respectively placed into the red blood cell suspension obtained in step S2 and incubated at 37°C for 1 hour. The PCL fiber membranes of each group were then removed, and the red blood cell suspensions were centrifuged at 3000g for 5 minutes. The supernatant was collected and its absorbance at 540nm was measured using an ELISA reader. PBS (0.01 mol·L⁻¹) was then added... -1 Red blood cell suspensions diluted with deionized water were used as negative control (NC) and positive control (PC), respectively.

[0161] Figure 9 The diagram shows the hemolysis test results of various groups of PCL fiber membranes with different surface coatings. The PCL fiber membrane with a PDA coating (corresponding to...) Figure 9 PCL-PDA group), PCL fiber membrane with PDA-2,3-DHTA COF composite coating on surface (corresponding to Figure 9 PCL-PDA-Cu group), PCL fiber membrane with PDA-Co-2,3-DHTA COF composite coating on surface (corresponding to Figure 9 After co-incubation with red blood cells, the PCL-PDA-CuCo group did not cause red blood cell rupture. The supernatant after co-incubation had low OD values, which were significantly lower than those of the positive control group and similar to those of the negative control group. This indicates that the PCL fiber membrane with a PDA-Co-2,3-DHTA COF composite coating of the present invention has excellent blood compatibility.

[0162] 6. Cobalt ion release tests were performed on the PCL fiber membranes with PDA-Co-2,3-DHTA COF composite coatings prepared in Example 2 (i.e., the 2,3-Dha COF group), the PCL fiber membranes with PDA-Co-2,5-DHTA COF composite coatings prepared in Comparative Example 3 (i.e., the 2,5-Dha COF group), and the PCL fiber membranes with PDA-Co-Bpy COF composite coatings prepared in Comparative Example 4 (i.e., the Bpy COF group). The test method was as follows: The PCL fiber membranes with different coatings were placed in a shaker at 37°C and incubated in 1 mL of 10% fetal bovine serum solution (pH = 7.4). At each specified time, the old solution was completely collected and replaced with a new solution. Then, the cobalt ion concentration in the solution was checked by inductively coupled plasma mass spectrometry (ICP-MS, Agilent 7700). A series of known Co... 2+ Create a standard curve concentration (in ddH). Figure 10 The figure shows the cobalt ion release of each group of PCL fiber membranes. Figure 10 It is known that the coordination mode of cobalt ions in copper tetraphenylporphyrin-cobalt covalent organic framework polymer materials has a crucial impact on the release rate of cobalt ions in vivo. The OO coordination mode of cobalt ions in the Co-2,3-DHTA COF of this invention is significantly superior to the NO coordination mode in Comparative Example 3 and the NN coordination mode in Comparative Example 4, endowing cobalt ions with more stable and controllable release behavior. Specifically, the release rate of cobalt ions can be effectively regulated, thereby maintaining it within an ideal biologically effective concentration range in the in vivo environment, avoiding the initial concentration surge caused by excessively rapid release or the ineffectiveness caused by excessively slow release, and promoting the recovery of HUVECs cell function and endogenous NO release. This advantage is particularly prominent in biomaterials and related applications, fully meeting the dual clinical needs for controlled release of metal ions and safety performance.

[0163] 7. The pore structure and specific surface area of ​​Co-2,3-DHTA COF prepared in Example 1, Co-2,5-DHTA COF prepared in Comparative Example 1, and Co-Bpy COF prepared in Comparative Example 2 were tested using the following methods:

[0164] (1) Sample pretreatment: Take an appropriate amount of COF material powder and put it into a clean sample tube. Accurately weigh the total mass of the sample tube and the sample (record the initial mass).

[0165] (2) Degassing treatment: Install the sample tube into the degassing station of the 3flex instrument, set the degassing temperature and time, and start the vacuum degassing program to ensure that the moisture on the material surface is fully removed. After degassing, weigh the sample tube and the sample again to calculate the actual sample mass.

[0166] (3) Sample installation: Transfer the degassed sample tube to the 3flex analysis station and connect it to the liquid nitrogen Dewar flask to ensure that the liquid nitrogen temperature (77K) is maintained throughout the experiment.

[0167] (4) Parameter settings: In the instrument software, select the "full-range adsorption-desorption" mode, set nitrogen as the adsorbed gas, and input parameters such as sample mass. After setting the relative pressure range, use coverage surface area (BET) and pore size distribution (DFT / BJH) to analyze the required data points.

[0168] (5) Data acquisition: Start the experiment and the instrument will automatically perform nitrogen adsorption-desorption cycle, record the adsorption amount data under different relative pressures, and generate a complete adsorption isotherm.

[0169] The pore size of the 2,3-DHTA COF prepared in Example 1 is 2.32 nm, and the pore size of the Co-2,3-DHTA COF prepared in Example 1 is 2.04 nm. The specific surface area test results for both are shown in [reference needed]. Figure 11 A.

[0170] The pore size of the 2,5-DHTA COF prepared in Comparative Example 1 was 2.29 nm, and the pore size of the Co-2,5-DHTA COF prepared in Comparative Example 1 was 1.92 nm. The specific surface area test results for both are shown in [reference needed]. Figure 11 B.

[0171] The pore size of the Bpy COF prepared in Comparative Example 2 was 3.15 nm, and the pore size of the Co-Bpy COF prepared in Comparative Example 2 was 2.98 nm. The specific surface area test results for both are shown in [reference needed]. Figure 11 C.

[0172] The above description is merely a specific implementation of this application. Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the specific working process described above can be referred to the corresponding process in the foregoing method embodiments, and will not be repeated here. It should be understood that the protection scope of this application is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in this application, and these modifications or substitutions should all be covered within the protection scope of this application.

Claims

1. A transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material, characterized in that, Including the repeating unit shown in equation (Ⅰ), Wherein, X is selected from Cu 2+ Ni 2+ Fe 2+ Zn 2+ and Co 2+ One of them; R1 and R2 are each independently selected from one of -H, alkyl, hydroxyl, halogen, aryl, cycloalkyl, hydroxyalkyl, alkoxy, thioalkoxy, nitro, cyano, and amino.

2. The transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material according to claim 1, characterized in that, The specific surface area of ​​the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material is 510 m². 2 ·g -1 -520m 2 ·g -1 ; and / or, The transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material includes multiple pores, the average pore size of which is 2.00 nm to 2.20 nm.

3. A method for preparing a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material as described in any one of claims 1-2, characterized in that, include: A covalent organic framework material intermediate was prepared by mixing 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal, 2,3-dihydroxyterephthalaldehyde, a first solvent, and a catalyst via a Schiff base condensation reaction. The covalent organic framework material intermediate was mixed with cobalt nitrate hexahydrate and a second solvent and subjected to a coordination reaction to obtain a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material.

4. The preparation method according to claim 3, characterized in that, The first solvent includes at least one selected from o-dichlorobenzene, m-dichlorobenzene, p-dichlorobenzene, toluene, xylene, n-butanol, methanol, ethanol, benzyl alcohol, phenylethanol, and phenylpropanol; and / or, The catalyst comprises at least one selected from acetic acid, formic acid, hydrochloric acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, and p-toluenesulfonic acid; and / or, The second solvent includes at least one of n-butanol, methanol, ethanol, and water.

5. The preparation method according to claim 3, characterized in that, The molar ratio of 5,10,15,20-tetra-(4-aminophenyl)-porphyrin-transition metal to 2,3-dihydroxyterephthalaldehyde is (0.4-0.6):1; and / or, The catalyst has a molar concentration of 5 mol / L to 7 mol / L; and / or, The mass ratio of the covalent organic framework material intermediate to cobalt nitrate hexahydrate is (0.04-0.06):1; and / or, The Schiff base condensation reaction is carried out at a temperature of 110°C-130°C; and / or, The Schiff base condensation reaction takes 60-80 hours; and / or, The coordination reaction is carried out at a temperature of 90℃-100℃; and / or, The reaction time for the coordination reaction is 20-30 hours.

6. A medical material, characterized in that, It includes functional components and matrix materials, wherein the functional components include the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material according to any one of claims 1-2 or the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method according to any one of claims 3-5.

7. The medical material according to claim 6, characterized in that, The medical materials include implantable materials or external contact materials; Preferably, the implantable materials include vascular implants and non-vascular implants; Preferably, the vascular implant includes at least one of an artificial blood vessel and a vascular stent; Preferably, the non-vascular implant includes at least one of a heart valve, a tissue-engineered scaffold, a catheter, and an in vivo implantable lead; Preferably, the in vitro contact material includes dialysis tubing.

8. The medical material according to claim 6, characterized in that, The medical material also includes a substrate material; and / or, The matrix material includes at least one of polydopamine, gelatin, chitosan, hyaluronic acid, and polyethyleneimine; and / or, The substrate material includes at least one of polymer materials, metals and alloys, ceramics and composites, medical glass, carbon-based materials, and bio-based materials.

9. The application of a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material according to any one of claims 1-2, or a transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material prepared by the preparation method according to any one of claims 3-5, in the preparation of biomedical materials for achieving complementary release of endogenous NO and exogenous NO, characterized in that, The biomedical materials mentioned include one or more of the following: (1) A medical coating material, wherein the medical coating material comprises the medical material according to claim 6; (2) A medical device, wherein the medical device includes the medical coating material; (3) Drug carrier; (4) Nanocatalysts; Preferably, the medical device includes implantable materials or materials that come into contact with the body.

10. The application according to claim 9, characterized in that, The method for preparing the medical device includes: The precursor raw material and the transition metal tetraphenylporphyrin-cobalt covalent organic framework polymer material were dissolved in a buffer solution to obtain a precursor solution; The precursor solution is coated onto the surface of a substrate material to obtain a medical device with a medical coating material on its surface; The precursor raw materials include at least one of dopamine, gelatin, chitosan, hyaluronic acid, and polyethyleneimine, and the coating method includes at least one of spraying, spin coating, dip coating, physical adsorption, chemical grafting, cross-linking curing, sol-gel method, blend coating method, and layer-by-layer self-assembly method.

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