A drug structure coating with long-acting immunomodulatory function and a preparation method thereof

Abstract

The application belongs to the technical field of biomedical functional materials, and particularly relates to a drug structure coating with long-acting immunoregulation function and a preparation method thereof. The preparation method is as follows: mixing a nano drug carrier suspension, a polyphenol compound acidic buffer, a polyamine compound acidic buffer and an oxidant acidic buffer, and reacting on the surface of a base material. The preparation method utilizes the oxidation, cross-linking and polymerization reaction of the polyphenol and polyamine compound under the action of the oxidant to generate a polyphenol nanoparticle (10 nanometer level) film layer outside the nano drug carrier (100 nanometer level). The polyphenol nano film layer not only serves as the "protective layer" of the nano drug carrier and the cross-linking material therebetween, but also can fix the nano drug carrier on the surface of the base material, and together with the drug carrier, constructs a drug structure coating with a topological structure. The hydrophilic groups contained in the coating make the surface super-hydrophilic and negatively charged, and make the coating have excellent biocompatibility.

Description

Technical Field

[0001] This invention belongs to the field of biomedical functional materials technology, specifically relating to a drug structure coating with long-lasting immunomodulatory function and its preparation method. Background Technology

[0002] Biomaterials are specialized functional materials capable of diagnosing, treating, replacing, repairing, inducing, regenerating, or enhancing the function of cells, tissues, and organs in the body. However, the implantation / mediation process of biomaterials is invasive, causing damage to the body and triggering inflammatory reactions. After implantation, the biomaterial, as a foreign body, interacts with body fluids, leading to protein adsorption on the biomaterial surface and initiating coagulation cascades, the complement system (polarizing immune cells into inflammatory responses), and the formation of temporary matrices. Furthermore, the chemical and physical properties of the material itself, along with in-situ movement or infection of the implant, cause continuous stimulation to the implantation site, leading to chronic inflammation, vascular and connective tissue proliferation; this can result in the implant being encapsulated by fibers, causing loss of implant function and various complications. Foreign body reactions cause patient suffering and significant economic losses, representing a clinical challenge and a bottleneck restricting the development of implantable materials and medical devices.

[0003] The three traditional strategies for combating foreign body reactions are:

[0004] 1) By altering the physical properties of the implanted material, such as its size, shape, hardness, and surface topology;

[0005] 2) Combine anti-inflammatory drugs or biological agents to suppress the inflammatory response;

[0006] 3) Modify the surface of implant materials with bioactive molecules to regulate foreign body reactions.

[0007] These traditional strategies have solved the problem of foreign body reaction to a certain extent and within a certain range; however, each of these strategies has its own shortcomings, as follows:

[0008] While optimizing the physical properties of materials can play a role in the first strategy, there is no established rule or experience regarding what physical properties can minimize foreign body reactions. Furthermore, different implant materials and devices require specific physical properties, so this method is only applicable under certain specific conditions and is not universally applicable.

[0009] The second strategy, which combines anti-inflammatory drugs (such as corticosteroids) or biologics (such as anti-inflammatory protein inhibitors), is a common approach to combat inflammation and foreign body reactions. However, these drugs or biologics raise concerns about toxicity and side effects. Furthermore, drug stability issues make it difficult to achieve long-term anti-foreign body reaction efficacy with this strategy.

[0010] The third strategy, bioactivity strategy, is also not a universal approach. For the diverse range of biomaterials, specific functional modifications are usually required on the surface of each implant or medical device for a particular purpose to meet the needs.

[0011] Furthermore, the root cause of foreign body reactions—the non-specific adsorption of proteins—remains difficult to overcome in traditional anti-foreign body reaction strategies.

[0012] Therefore, based on this, the technical solution of the present invention is proposed. Summary of the Invention

[0013] To address the problems existing in the prior art, this invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, comprising the following steps:

[0014] (1) Disperse the nanomedicine carrier in a solvent to obtain a nanomedicine carrier suspension;

[0015] (2) The nanomedicine carrier suspension is mixed with polyphenol compound acid buffer, polyamine compound acid buffer and oxidant acid buffer to obtain a mixture;

[0016] (3) The mixture is added to the surface of the substrate material to react. After the reaction is completed, it is cleaned and dried to obtain a drug structure coating with long-term immunomodulatory function.

[0017] To facilitate understanding of this invention, the reaction principle of this invention will be explained as follows:

[0018] This invention first uniformly disperses a nanocarrier loaded with an anti-inflammatory drug in a solvent. Under acidic conditions, polyphenolic and polyamine compounds are oxidized, cross-linked, and polymerized in the presence of an oxidizing agent to generate a film layer with a polyphenol nanoparticle structure (tens of nanometers). This nanofilm layer uniformly encapsulates the nano-drug carrier (hundreds of nanometers) and firmly fixes it to the surface of a substrate material, forming a topological structure and constructing a drug-structured coating with the drug as the bulk material. The topological structure and numerous hydrophilic groups in the coating interact to give the surface superhydrophilicity and a negative charge. Ultimately, the coating exerts different biological functions at different stages after device implantation through a superhydrophilic antifouling platform, the synergistic effect of polyphenols and anti-inflammatory drugs, and long-term regulation of the immune response in the lesion microenvironment. This technology can be used for surface modification of long-term implantable devices / materials such as catheters, bone implants, cardiovascular materials, and ophthalmic materials.

[0019] Preferably, in step (1), the drug is one or a combination of two or more of the following: curcumin, triptolide, triptolide, astragaloside A, astragalus polysaccharide, baicalin, magnolol, honokiol, ginsenosides, lutein, flavonoids, quercetin, interferon, and interleukin-2.

[0020] Preferably, in step (1), the carrier is one of exosomes, solid lipid nanoparticles, nanoliposomes or polymer micelles.

[0021] Preferably, in step (1), the solvent is an aqueous solution, an ethanol solution, or a mixture of water and ethanol; in the mixture of water and ethanol, the volume ratio of ethanol to water is 0.1-5:0.1-5.

[0022] Preferably, in step (2), the polyphenolic compound is one or a combination of two or more of the following: tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol, and flavonoids; the polyamine compound is one or a combination of two or more of the following: ethylenediamine, pentanediamine, 2,2,4-trimethylhexamethylenediamine, 1,8-diaminooctane, methylcyclohexanediamine, 1,3-diaminomethylcyclohexane, 2,4,6-triaminomethylcyclohexane, 1,4-bis(diaminohexylcyclohexane), and 2-(3,4-dihydroxyphenyl)ethylamine; and the oxidant is one or a combination of two or more of the following: hydrogen peroxide, ammonium persulfate, concentrated nitric acid, sodium periodate, potassium permanganate, and potassium dichromate.

[0023] Preferably, in step (2), the pH of the acidic buffer solution is 3 to 7; the acidic buffer solution is one of the following: acetic acid-acetate buffer solution, 2-(N-morpholino)ethanesulfonic acid buffer solution, glycine-hydrochloric acid buffer solution, phthalic acid-hydrochloric acid buffer solution, potassium hydrogen phthalate-sodium hydroxide buffer solution, disodium hydrogen phosphate-citric acid buffer solution, citric acid-sodium hydroxide-hydrochloric acid buffer solution, and citric acid-sodium citrate buffer solution.

[0024] Preferably, in step (2), the concentration of the nanomedicine carrier in the mixture is 0.5–10 mg / mL, the concentration of the polyphenol compound is 0.5–20 mg / mL, the concentration of the polyamine compound is 0.1–10 mg / mL, and the concentration of the oxidant is 0.1–10 mg / mL. More preferably, the concentration of the nanomedicine carrier is 2 mg / mL, the concentration of the polyphenol compound is 1 mg / mL, the concentration of the polyamine compound is 0.5 mg / mL, and the concentration of the oxidant is 0.5 mg / mL.

[0025] Preferably, in step (3), the substrate material is one of metal-based biomaterials, polymer-based biomaterials, ceramic-based biomaterials, and composite biomaterials.

[0026] Preferably, in step (3), the reaction temperature is 10–40°C and the reaction time is 0.1–10 h. More preferably, the reaction temperature is 30°C and the reaction time is 1 h.

[0027] Based on the same technical concept, another aspect of the present invention is to provide a drug structure coating with long-lasting immunomodulatory function obtained by the above preparation method.

[0028] The beneficial effects of this invention are as follows:

[0029] 1. The preparation method of the present invention is based on the oxidation, cross-linking, polymerization and deposition reactions of polyphenols and polyamine compounds under the action of oxidants. A uniform polyphenol nanoparticle film layer can be formed on the surface of the nanomedicine carrier using a simple "one-pot" method. The polyphenol nanoparticle film layer not only increases the stability of the nanomedicine carrier, but also firmly fixes the nanomedicine carrier to the surface of the substrate material. It realizes the bulk material constructed with the drug as a coating. As long as the coating exists, the drug exists and can exert biological functions for a long time. Moreover, the polyphenol nanoparticle film layer is non-material dependent and can be modified and modified on the surface of substrate materials with different shapes and properties, and has wide applicability.

[0030] 2. Unlike traditional methods for preparing topological structures, the method described in this invention produces a drug structure coating with polyphenol nanoparticles in the tens of nanometers range and a nano-drug carrier in the hundreds of nanometers range. The combination of nanoparticles of different sizes gives the coating a topological structure, which possesses bioactivity and biological function without damaging the substrate. Simultaneously, the coating contains numerous hydrophilic groups such as phenolic hydroxyl, carboxyl, and amino groups, which, together with the topological structure, impart excellent hydrophilicity to the drug structure coating. Furthermore, the coating surface is negatively charged, giving it excellent resistance to non-specific protein adhesion, thus preventing acute thrombosis and acute inflammation. The polyphenol nanofilm also possesses the corresponding biological functions of polyphenols, such as anti-oxidation, free radical scavenging, inflammation regulation, and vascular ECs-friendly properties.

[0031] 3. Unlike traditional drug-loaded coatings, the coating constructed by the preparation method of the present invention is a bulk material constructed with the drug as the coating. As long as the coating exists, the drug exists and can exert the corresponding biological function. Moreover, the outer layer of the nano-drug carrier is wrapped by a polyphenol nanocomposite layer, which can achieve controllable and long-term drug release.

[0032] 4. Unlike traditional methods that regulate the biological properties of materials by a single factor, the drug-structured coating constructed by the preparation method described in this invention regulates the inflammatory response of the lesion site microenvironment through multiple factors (such as a superhydrophilic platform, polyphenols, and anti-inflammatory drugs) in a synergistic and non-interfering preferred mode, providing favorable conditions for the recovery of vascular tissue.

[0033] 5. The preparation method described in this invention is simple to operate, does not require large-scale equipment, has high reaction efficiency, mild conditions, and has broad applicability. Attached Figure Description

[0034] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0035] Figure 1 The image shows a scanning electron microscope (SEM) image of the surface morphology of the drug structure coating with long-acting immunomodulatory function prepared in Example 1.

[0036] Figure 2 The surface water contact angle (WCA) of the drug structure coating with long-lasting immunomodulatory function prepared in Example 1;

[0037] Figure 3 The fluorescence intensity of the drug structure coating with long-acting immunomodulatory function prepared in Example 1 is measured by the fluorescence intensity of bovine serum albumin (FITC-BSA) and human fibrinogen (FITC-HFg) labeled with fluorescein isothiocyanate on the surface of the drug structure coating.

[0038] Figure 4 The image shows the cell morphology of the drug structure coating with long-acting immunomodulatory function prepared in Example 1, which inhibits macrophage adhesion and growth. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0040] Example 1

[0041] This invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, the method comprising the following steps:

[0042] (1) Tween-80 micelles (polymer micelles) loaded with magnolol were uniformly dispersed in a mixture of ethanol and water to obtain a nano-drug carrier suspension; wherein the volume ratio of ethanol to water was 1:4.

[0043] (2) Tannic acid, ethylenediamine and sodium periodate were dissolved in sodium acetate buffer solution with pH=5 to obtain tannic acid buffer solution, ethylenediamine buffer solution and sodium periodate buffer solution respectively;

[0044] (3) The nanomedicine carrier suspension, the tannic acid buffer, the ethylenediamine buffer and the sodium periodate buffer are mixed to obtain a mixture; wherein the concentrations of drug-loaded micelles, tannic acid, ethylenediamine and sodium periodate are 2 mg / mL, 1 mg / mL, 0.5 mg / mL and 0.5 mg / mL, respectively.

[0045] (4) Add the mixture to the clean surface of the polymer substrate, then react at 25°C for 3 hours, thoroughly clean with RO water, and dry with nitrogen to obtain a drug structure coating with long-lasting immunomodulatory function.

[0046] Example 2

[0047] This invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, the method comprising the following steps:

[0048] (1) The lutein-loaded nanoliposomes were uniformly dispersed in water to obtain a nanomedicine carrier suspension.

[0049] (2) Epigallocatechin gallate, pentanediamine and potassium permanganate were dissolved in sodium acetate buffer at pH=4 to obtain epigallocatechin gallate buffer, pentanediamine buffer and potassium permanganate buffer.

[0050] (3) The nanomedicine carrier suspension, the epigallocatechin gallate buffer, the pentanediamine buffer and the potassium permanganate buffer are mixed to obtain a mixture; wherein the concentrations of the drug-loaded nanoliposomes, epigallocatechin gallate, pentanediamine and potassium permanganate are 5 mg / mL, 2 mg / mL, 2 mg / mL and 2 mg / mL, respectively.

[0051] (4) Add the mixture to the surface of a clean metal substrate material, then react at 20°C for 6 hours, thoroughly clean with RO water, and dry with nitrogen to obtain a drug structure coating with long-lasting immunomodulatory function.

[0052] Example 3

[0053] This invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, the method comprising the following steps:

[0054] (1) Lecithin loaded with baicalin was uniformly dispersed in water to obtain a nano-drug carrier suspension;

[0055] (2) Gallic acid, 2,2,4-trimethylhexamethylenediamine and sodium periodate were dissolved in phthalic acid-hydrochloric acid buffer solution with pH=3.5 to obtain gallic acid buffer solution, 2,2,4-trimethylhexamethylenediamine buffer solution and sodium periodate buffer solution.

[0056] (3) The nanomedicine carrier suspension, the gallic acid buffer, the 2,2,4-trimethylhexamethylenediamine buffer and the sodium periodate buffer are mixed to obtain a mixture; wherein the concentrations of drug-loaded lecithin, gallic acid, 2,2,4-trimethylhexamethylenediamine and sodium periodate are 6 mg / mL, 3 mg / mL, 3 mg / mL and 3 mg / mL, respectively;

[0057] (4) Add the mixture to the surface of a clean metal substrate material, then react at 15°C for 10 hours, thoroughly clean with RO water, and dry with nitrogen to obtain a drug structure coating with long-lasting immunomodulatory function.

[0058] Example 4

[0059] This invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, the method comprising the following steps:

[0060] (1) The triptolide exosomes were uniformly dispersed in water to obtain a nano-drug carrier suspension;

[0061] (2) Dissolve pyrogallol, 1,8-diaminooctane and ammonium persulfate in citrate-sodium citrate buffer solution at pH=4 to obtain pyrogallol buffer solution, 1,8-diaminooctane buffer solution and ammonium persulfate buffer solution respectively.

[0062] (3) The nanomedicine carrier suspension, the pyrogallol buffer, the 1,8-diaminooctane buffer and the ammonium persulfate buffer are mixed to obtain a mixture; wherein the concentrations of the drug-loaded exosomes, pyrogallol, 1,8-diaminooctane and ammonium persulfate are 2 mg / mL, 2 mg / mL, 1.5 mg / mL and 2 mg / mL, respectively.

[0063] (4) Add the mixture to the surface of a clean metal substrate material, react at 35°C for 1 hour, thoroughly clean with RO water, and dry with nitrogen to obtain a drug structure coating with long-lasting immunomodulatory function.

[0064] Example 5

[0065] This invention provides a method for preparing a drug structure coating with long-lasting immunomodulatory function, the method comprising the following steps:

[0066] (1) The nanoliposomes loaded with magnolol were uniformly dispersed in water to obtain a nanodrug carrier suspension.

[0067] (2) The epicatechin gallate, 1,3-diaminomethylcyclohexane and potassium dichromate were dissolved in glycine-hydrochloric acid buffer solution at pH=6 to obtain epicatechin gallate buffer solution, 1,3-diaminomethylcyclohexane buffer solution and potassium dichromate buffer solution respectively.

[0068] (3) The nanomedicine carrier suspension, the epicatechin gallate buffer, the 1,3-diaminomethylcyclohexane buffer and the potassium dichromate buffer are mixed to obtain a mixture; wherein the concentrations of the drug-loaded nanoliposomes, epicatechin gallate, 1,3-diaminomethylcyclohexane and potassium dichromate are 6 mg / mL, 2 mg / mL, 2 mg / mL and 2 mg / mL, respectively;

[0069] (4) Add the mixture to the surface of a clean metal substrate material, react at 35°C for 0.5 h, thoroughly clean with RO water, and dry with nitrogen to obtain a drug structure coating with long-lasting immunomodulatory function.

[0070] Test case

[0071] SEM images of the drug structure coating with long-lasting immunomodulatory function obtained in Example 1 are shown below. Figure 1 As shown in the figure, a multi-level structure coating with nano-drug carriers as the main body is generated on the surface of the substrate material. The outer surface of the nano-drug carriers in the coating is uniformly covered by polyphenol nanoparticles with a particle size of 10-20 nm. Moreover, the reaction process does not change the morphology of the nano-drug carriers, which have a multi-level structure.

[0072] from Figure 2 The water contact angle results show that the water contact angle of the drug-grade multi-level structure coating is less than 5°, indicating superhydrophilicity. (Where Blank is the blank control group, and Coating is the experimental group coated with the drug-grade structure coating, the same applies below.)

[0073] from Figure 3 After fluorescently labeled human fibrinogen was adsorbed onto drug-coated and unmodified plastic substrates, the fluorescence intensity of the adsorbed proteins was measured; the higher the intensity, the greater the amount of adsorbed protein. The hydration layer formed on the superhydrophilic surface upon contact with the liquid effectively resists protein adhesion.

[0074] from Figure 4 It can be observed that, under the synergistic effect of the superhydrophilic platform and polyphenol drugs, the surface of the drug multi-level structure coating can effectively resist the adhesion and activation of macrophages.

[0075] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing a drug structure coating having a long-acting immunomodulatory function, characterized by, The method comprises the following steps: (1) dispersing the nano-drug carrier in a solvent to obtain a nano-drug carrier suspension; (2) mixing the nano-drug carrier suspension with a polyphenol compound acid buffer, a polyamine compound acid buffer and an oxidizing agent acid buffer to obtain a mixed solution; (3) adding the mixed solution to the surface of the base material for reaction, and then cleaning and drying to obtain a drug structure coating with long-term immunomodulatory function; wherein: In an acidic environment, the polyphenol compound and the polyamine compound are oxidized, cross-linked and polymerized under the action of the oxidizing agent to form a film layer with a polyphenol nanoparticle structure, and the size of the polyphenol nanoparticle structure is ten nanometers; the film layer with the polyphenol nanoparticle structure uniformly wraps and firmly fixes the nano-drug carrier on the surface of the base material, forms a topological structure, and constructs a drug structure coating with the drug as the coating bulk material, and the size of the nano-drug carrier is hundreds of nanometers.

2. The method for preparing the drug structure coating with long-acting immunomodulatory function according to claim 1, characterized in that, In step (1), the drug is one or a combination of two or more of curcumin, triptolide, tripterine, astragaloside, astragalus polysaccharide, baicalin, honokiol, magnolol, ginsenoside, lutein, flavonoid quercetin, interferon and interleukin 2.

3. The method for preparing the drug structure coating with long-lasting immunomodulatory function according to claim 1, characterized in that, In step (1), the carrier is one of exosomes, solid lipid nanoparticles, nanoliposomes or polymer micelles.

4. The method for preparing the drug structure coating with long-acting immunomodulatory function according to claim 1, characterized in that, In step (1), the solvent is an aqueous solution, an ethanol solution or a mixed solution of water and ethanol; in the mixed solution of water and ethanol, the volume ratio of ethanol to water is 0.1-5:0.1-5.

5. The method for preparing the drug structure coating with long-acting immunomodulatory function according to claim 1, characterized in that, In step (2), the polyphenol compound is one or a combination of two or more of tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol and flavones; the polyamine compound is one or a combination of two or more of ethylenediamine, pentanediamine, 2,2,4-trimethylhexamethylene diamine, 1,8-diamino octane, methylcyclohexane diamine, 1,3-diaminomethylcyclohexane, 2,4,6-triaminomethylcyclohexane, 1,4-bis-diaminohexylcyclohexane and 2-(3,4-dihydroxyphenyl)ethylamine; and the oxidizing agent is one or a combination of two or more of hydrogen peroxide, ammonium persulfate, concentrated nitric acid, sodium periodate, potassium permanganate and potassium dichromate.

6. The method for preparing the drug structure coating with long-lasting immunomodulatory function according to claim 1, characterized in that, In step (2), the pH of the acid buffer is 3-7; and the acid buffer is one of acetic acid-acetate buffer, 2-(N-morpholine)ethanesulfonic acid buffer, glycine-hydrochloric acid buffer, phthalic acid-hydrochloric acid buffer, potassium hydrogen phthalate-sodium hydroxide buffer, disodium hydrogen phosphate-citric acid buffer, citric acid-sodium hydroxide-hydrochloric acid buffer and citric acid-sodium citrate buffer.

7. The method for preparing the drug structure coating with long-acting immunomodulatory function according to claim 1, characterized in that, In step (2), in the mixed solution, the concentration of the nano-drug carrier is 0.5-10 mg / mL, the concentration of the polyphenol compound is 0.5-20 mg / mL, the concentration of the polyamine compound is 0.1-10 mg / mL, and the concentration of the oxidizing agent is 0.1-10 mg / mL.

8. The method for preparing the drug structure coating with long-acting immunomodulatory function according to claim 1, characterized in that, In step (3), the base material is one of a metal-based biomaterial, a polymer-based biomaterial, a ceramic-based biomaterial, and a composite biomaterial.

9. The method of claim 1, wherein the drug structure coating having long- term immunomodulatory function is prepared by the steps of: (a) preparing a solution of the drug structure coating having long-term immunomodulatory function; (b) coating the solution on a substrate; (c) drying the solution; and (d) removing the substrate. In step (3), the reaction temperature is 10-40℃, and the reaction time is 0.1-10h.

10. The drug structure coating with long-acting immunomodulatory function obtained by the preparation method in any one of claims 1-9.

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