Preparation method of anticoagulant blood contact material

Abstract

The invention discloses a preparation method of an anticoagulant blood contact material, which comprises the following steps: S1, dissolving a silane reagent containing different functional groups by using ethanol, immersing the blood contact material into the silane solution, and reacting for a period of time in a constant-temperature environment in a dark place for later use; s2, immersing the sample obtained in the step S1 in an ethanol solution containing 1mM of TCEP (tris (2-carboxyethyl) phosphine), reacting, fully cleaning, and carrying out vacuum drying for later use; s3, dissolving an anticoagulant with a PBS (Phosphate Buffer Solution) to obtain an anticoagulant solution for later use; and S4, immersing the sample obtained in the step S2 in the anticoagulant solution prepared in the step S3, fully reacting at a constant temperature, cleaning, and drying with nitrogen to obtain the anticoagulant blood contact material. The method is simple to operate, mild in reaction condition, green, economical, free of large equipment and convenient to popularize and use. The silane reagent is used for treating the blood contact material, so that active groups such as sulfydryl, carboxyl, amido and hydroxyl can be generated on the surface of the material in situ, and meanwhile, the hydrophilicity of the material is improved. Then, the functionalized material is immersed in an anticoagulant solution, an anticoagulant drug is fixed on the surface of the material through a mild click chemical reaction, an amidation reaction and the like, and the blood contact material is endowed with excellent anticoagulant performance.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical functional materials, and in particular relates to a method for preparing an anticoagulant blood-contact material, which can be used for surface modification of blood-contact materials and devices. Background Art

[0002] When blood-contact medical devices such as catheters (disposable catheters, central venous catheters, hemodialysis circulation catheters, etc.) and vascular stents (arterial stents, venous stents), etc., come into contact with body tissues or blood, proteins will undergo non-specific adhesion and denaturation on the material surface. Adherent and denatured proteins will further promote the adhesion of cells and bacteria, triggering acute thrombosis and acute inflammation, thereby leading to device implantation failure. In addition, a large amount of adhered proteins will cover the bioactive components on the surface of the device / material, affecting its biological function. Therefore, resisting protein adhesion and denaturation at the source is crucial to improving the biological function of the material. In the field of anti-protein adhesion, hydrophilic surfaces have been widely studied, mainly including polyethylene glycol coatings and amphiphilic ion molecular brush surfaces. These two types of coatings can effectively resist the adhesion of proteins and cells by effectively combining with water molecules to form a hydration layer on the surface. However, the anti-fouling performance of such coatings is highly dependent on the length and density of the modified molecular chains, and long-term use may lead to immune inflammation and other problems. Therefore, it is crucial to construct a new type of superhydrophilic surface.

[0003] As the service time in the body increases, its hydration layer will eventually be destroyed. Once destroyed, it will no longer be able to effectively maintain its long-term anti-fouling performance. Therefore, it is necessary to use drugs to exert corresponding biological functions. However, most drug-loaded coatings have problems with timeliness and stability. Once the drug in the coating is completely released, the coating will no longer have the corresponding biological function. Therefore, it is urgent to develop a simple, efficient, green, economical and widely applicable method to design a drug-based coating material with super-hydrophilic anti-fouling properties. As long as the coating exists, the drug structure coating has a vital role in maintaining long-term anticoagulation and anti-inflammatory effects on the surface. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for preparing an anticoagulant blood contact material to address the problems of equipment limitations, complex synthesis process, insufficient drug loading in the coating, and short timeliness in the above-mentioned prior art.

[0005] The technical solution adopted in the present invention is as follows:

[0006] A method for preparing an anticoagulant blood contact material comprises the following steps:

[0007] S1. Dissolve silane reagents containing different functional groups in ethanol, immerse the blood-contact material in the silane solution, and react in a constant temperature environment in the dark for a period of time.

[0008] S2. The sample obtained in step S1 was immersed in an ethanol solution containing 1 mM TCEP (tris(2-carboxyethyl)phosphine) or a 1 mg / mL aqueous solution of EDC, and after the reaction, it was thoroughly washed, vacuum-dried, and set aside;

[0009] S3. Dissolve the anticoagulant with PBS solution to obtain an anticoagulant solution for later use;

[0010] S4. Immerse the sample obtained in step S2 in the anticoagulant solution prepared in step S3, allow to react fully at a constant temperature, wash, and dry with nitrogen to obtain an anticoagulant blood contact material.

[0011] The present invention first treats blood-contact materials with a silane reagent to generate reactive groups (such as thiol, carboxyl, amine, and hydroxyl) on the material surface in situ, simultaneously increasing the material's hydrophilicity. Subsequently, the functionalized material is immersed in an anticoagulant solution, where the anticoagulant drug is immobilized on the material surface through a mild click chemistry reaction, an amidation reaction, or other methods, imparting excellent anticoagulant properties to the blood-contact material. This method is simple to operate, operates under mild reaction conditions, is environmentally friendly, and is economical. It does not require large-scale equipment, making it readily adaptable for widespread use.

[0012] Furthermore, the S1 contains silane reagents with different functional groups: (3-mercaptopropyl)trimethoxysilane, (3-mercaptopropyl)triethoxysilane, (3-mercaptopropyl)triethoxysilane, (3-aminopropyl)triethoxysilane, and 3-carboxypropyltrimethoxysilane.

[0013] Furthermore, the blood contact materials in S1 are: activated carbon, cationic resin, anionic resin, copper-like membrane, cellulose acetate, polysulfone, polyethersulfone, polyacrylonitrile, polyvinyl alcohol, and polyurethane.

[0014] Furthermore, the anticoagulant in S3 is at least one of heparin, low molecular weight heparin, hirudin, argatroban, bivalirudin, fondaparinux sodium, citrate, aspirin, and nafamostat mesylate.

[0015] Furthermore, the final concentration of the silane reagent in S1 is 0.5-10 mg / mL, the reaction time is 1-48 h, and the reaction temperature is 15-50°C.

[0016] Furthermore, the final concentration of the anticoagulant in S3 is 1 to 20 mg / mL.

[0017] Furthermore, the reaction time in S4 is 1 to 48 hours, and the reaction temperature is 15 to 50°C.

[0018] The above method is used to prepare an anticoagulant blood contact material.

[0019] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0020] 1. The present invention is based on silanization treatment to efficiently form active hydrophilic groups, such as sulfhydryl, amine, carboxyl and hydroxyl groups, on the surface of blood-contact materials in situ, providing active sites for subsequent modification of anticoagulant drugs;

[0021] 2. Silanization treatment improves the wettability of the material surface and further improves its biocompatibility;

[0022] 3. Different from traditional drug-loaded coatings, the coating constructed by this method fixes anticoagulant drugs on the surface of the material through mild reactions such as click chemistry and amidation reactions, which greatly improves the loading capacity and stability of anticoagulant drugs on the surface and effectively extends the service life of the material or device.

[0023] 4. The present invention has simple operation, efficient reaction, mild conditions and broad spectrum practicality. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0025] Figure 1 The surface heparin content of the anticoagulant coating prepared in Example 1;

[0026] Figure 2 is the surface water contact angle value of the anti-coagulation coating prepared in Example 2.

[0027] Figure 3 This is a scanning electron micrograph of platelets adhered to the surface of the anticoagulant coating prepared in Example 3. DETAILED DESCRIPTION

[0028] In order to make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only intended to explain the present invention and are not intended to limit the present invention. That is, the embodiments described herein are only some embodiments of the present invention, not all embodiments. Generally, the components of the embodiments of the present invention described and illustrated in the drawings herein may be arranged and designed in various different configurations.

[0029] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention as claimed, but is merely intended to represent selected embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative work are within the scope of protection of the present invention.

[0030] It should be noted that relational terms such as "first" and "second" are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of additional identical elements in the process, method, article, or apparatus comprising the element.

[0031] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0032] Example 1

[0033] A preferred embodiment of the present invention provides a method for preparing an anticoagulant blood contact material, the specific steps of which are as follows:

[0034] S1. Dissolve (3-aminopropyl)triethoxysilane containing an amino functional group in ethanol, immerse the polyethersulfone membrane in the blood contact material in the (3-aminopropyl)triethoxysilane solution, and react in a dark environment at 25°C for 2 hours before use;

[0035] S2. The sample obtained in step S1 was immersed in an aqueous solution containing 1 mg / mL EDC, reacted at 25°C for 2h, and then fully washed, vacuum-dried, and set aside;

[0036] S3. Dissolve heparin in PBS solution to obtain a heparin solution with a mass concentration of 4 mg / mL and set aside;

[0037] S4. Immerse the sample obtained in step S2 in the heparin solution prepared in step S3, react in an environment of 25°C for 2 hours, then fully wash and dry with nitrogen to obtain an anticoagulant blood contact material.

[0038] Example 2

[0039] A preferred embodiment of the present invention provides a method for preparing an anticoagulant blood contact material, the specific steps of which are as follows:

[0040] S1. Dissolve (3-mercaptopropyl)trimethoxysilane containing a thiol functional group in ethanol, immerse the polyurethane sheet in the blood contact material in the (3-mercaptopropyl)trimethoxysilane solution, and react in a dark environment at 20°C for 6 hours before use;

[0041] S2. The sample obtained in step S1 was immersed in an ethanol solution containing 1 mM TCEP (tris(2-carboxyethyl)phosphine), reacted at 20°C for 2 h, and then thoroughly washed, vacuum-dried, and set aside;

[0042] S3. Dissolve citrate in PBS solution to obtain a citrate solution with a concentration of 10 mg / mL and set aside;

[0043] S4. Immerse the sample obtained in step S2 in the heparin solution prepared in step S3, react in an environment of 25°C for 2 hours, then fully wash and dry with nitrogen to obtain an anticoagulant blood contact material.

[0044] Example 3

[0045] A preferred embodiment of the present invention provides a method for preparing an anticoagulant blood contact material, the specific steps of which are as follows:

[0046] S1. Dissolve (3-mercaptopropyl)triethoxysilane containing a thiol functional group in ethanol, immerse the polyurethane sheet in the blood contact material in the (3-mercaptopropyl)triethoxysilane solution, and react in a dark environment at 30°C for 10 hours before use;

[0047] S2. The sample obtained in step S1 was immersed in an ethanol solution containing 1 mM TCEP (tris(2-carboxyethyl)phosphine), reacted at 30°C for 5 h, and then thoroughly washed, vacuum-dried, and set aside;

[0048] S3. Dissolve nafamostat mesylate with PBS solution to obtain a concentration of 10 mg / mL nafamostat mesylate solution for later use;

[0049] S4. Immerse the sample obtained in step S2 in the nafamostat mesylate solution prepared in step S3, react in an environment of 30°C for 1 hour, then fully wash and dry with nitrogen to obtain an anticoagulant blood contact material.

[0050] Experimental example

[0051] The heparin content of the product obtained in Example 1 is 3.5ug / cm 2, while the surface of the modified polyethersulfone membrane does not contain heparin, indicating that heparin has been successfully modified to the surface of the material, and the modified heparin content is within the effective range for anticoagulation. The water contact angle of the product obtained in Example 2 is 24±2°, while the water contact angle of the unmodified polyurethane surface is 73±3°, indicating that the anticoagulant coating also significantly improves the wettability of the blood-contacting material. The blood compatibility test of the product obtained in Example 3 was conducted. Through the platelet adhesion experiment, it was found that a large number of platelets adhered to the surface of the unmodified polyurethane, and the adhered platelets were in an activated and spread state; while only a very small number of platelets adhered to the surface of the polyurethane modified with nafamostat mesylate, and the adhered platelets were in a round resting state, indicating that the nafamostat mesylate coating has excellent blood compatibility.

[0052] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing an anticoagulant blood contact material, characterized in that: The following steps are involved: S1. Dissolve silane reagents containing different functional groups in ethanol, immerse the blood-contact material in the silane solution, and react in a constant temperature environment in the dark for a period of time. S2. The sample obtained in step S1 was immersed in an ethanol solution containing 1 mM TCEP (tris(2-carboxyethyl)phosphine) or a 1 mg / mL aqueous solution of EDC, and after the reaction, it was thoroughly washed, vacuum-dried, and set aside; S3. Dissolve the anticoagulant with PBS solution to obtain an anticoagulant solution for later use; S4. Immerse the sample obtained in step S2 in the anticoagulant solution prepared in step S3, allow to react fully at a constant temperature, wash, and dry with nitrogen to obtain an anticoagulant blood contact material.

2. The method for preparing the anticoagulant blood contact material according to claim 1, characterized in that: The silane reagents containing different functional groups in S1 include: silane reagents containing mercapto (-SH), (3-mercaptopropyl)trimethoxysilane, (3-mercaptopropyl)triethoxysilane, (3-mercaptopropyl)triethoxysilane, (3-aminopropyl)triethoxysilane, and 3-carboxypropyltrimethoxysilane.

3. The method for preparing the anticoagulant blood contact material according to claim 1, wherein: The blood contact materials in S1 are: activated carbon, cationic resin, anionic resin, copper-like membrane, cellulose acetate, polysulfone, polyethersulfone, polyacrylonitrile, polyvinyl alcohol, and polyurethane.

4. The method for preparing the anticoagulant blood contact material according to claim 1, wherein: The anticoagulant in S3 is at least one of heparin, low molecular weight heparin, hirudin, argatroban, bivalirudin, fondaparinux sodium, citrate, aspirin, and nafamostat mesylate.

5. The method for preparing the anticoagulant blood contact material according to claim 1, wherein: The final concentration of the silane reagent in S1 is 0.5-10 mg / mL, the reaction time is 1-48 h, and the reaction temperature is 15-50° C.

6. The method for preparing the anticoagulant blood contact material according to claim 1, characterized in that: The final concentration of the anticoagulant in S3 is 1-20 mg / mL.

7. The method for preparing the anticoagulant blood contact material according to claim 1, characterized in that: The reaction time in S4 is 1 to 48 hours, and the reaction temperature is 15 to 50°C.

8. A blood contact material with excellent anticoagulant performance obtained by the preparation method according to any one of claims 1 to 8.

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