Preparation method of anticoagulant coating and medical instrument

Abstract

The invention discloses a preparation method of an anti-coagulation coating and a medical instrument with the anti-coagulation coating. The preparation method comprises the following steps: coating a substrate subjected to hydroxylation treatment with an initiator coating to obtain an activated substrate; placing the activated substrate, a polymeric monomer and an auxiliary agent in a reaction vessel, and adjusting the interior of the reaction vessel to a water-free and oxygen-free environment in one or more protection operations; and placing the reaction container under a target reaction condition to react, so that the polymeric monomer is subjected to in-situ polymerization on the activated substrate, and the anticoagulant coating is obtained. The invention discloses a method for preparing a nano-scale anticoagulant coating by directly carrying out polymer in-situ polymerization on the surface of a substrate. The grafting density of a polymer on the surface of a material is improved, so that the anticoagulant effect of the anticoagulant coating is improved.

Description

Technical Field

[0001] The present application relates to the field of medical materials, and in particular to a method for preparing an anticoagulant coating and a medical device having the anticoagulant coating. Background Art

[0002] Implantable and interventional medical devices play an important role in medical processes such as disease diagnosis, treatment, monitoring or blood processing. However, these medical devices still face many challenges in clinical use, such as thrombosis, infection risk, immune response, etc., which have a great impact on medical effects. At present, one solution is to construct an anticoagulant coating on the surface of these medical devices. The anticoagulant coating can effectively reduce platelet adhesion and activation, while reducing fibrinogen adsorption, and inhibiting the activation of coagulation factors to prevent blood coagulation and avoid thrombosis. In addition, foreign body reactions and inflammatory reactions can also be effectively inhibited to improve the biocompatibility of medical devices, thereby increasing the safety and reliability of the devices. Therefore, it is very important to construct an anticoagulant coating with efficient anticoagulation function on the surface of implantable and interventional medical devices.

[0003] The existing method for preparing nano-scale anticoagulant coatings generally involves first coating a polymer primer with reactive functional groups on the surface of the device, and then grafting the polymer material used for anticoagulation onto the primer through the reaction of the terminal active group with the aforementioned reactive functional group. This method has two disadvantages. First, due to the steric hindrance effect of the polymer material, the grafting efficiency between the polymer material and the polymer material used for anticoagulation is low, which reduces the effect of the final anticoagulant coating. At the same time, the unreacted functional groups on the primer will affect the anticoagulant effect of the coating and the safety of the final product. Second, using a polymer as a primer will increase the thickness of the entire coating, which may affect the final device performance. Summary of the invention

[0004] In order to solve the above problems, the present application discloses a method for preparing an anticoagulant coating and a medical device having the anticoagulant coating. The preparation method directly grafts a polymer monomer for anticoagulation on the surface of a substrate to obtain an anticoagulant coating, thereby achieving a high grafting density of the anticoagulant material and improving the anticoagulant effect.

[0005] On the one hand, the present application discloses a method for preparing an anticoagulant coating, which comprises: coating an initiator coating on a hydroxylated substrate to obtain an activated substrate; placing the activated substrate, a polymerizable monomer, and an auxiliary agent in a reaction container, and adjusting the interior of the reaction container to an anhydrous and oxygen-free environment during one or more protective operations; placing the reaction container under target reaction conditions to react so that the polymerizable monomer is polymerized in situ on the activated substrate, thereby obtaining the anticoagulant coating.

[0006] According to some embodiments of the present application, obtaining the activated substrate includes: providing an initiator and preparing an initiator solution; the initiator is selected from compounds having methoxysilane and / or thiol; adding a catalyst to the initiator solution to obtain a mixed solution, and placing the hydroxylated substrate in the mixed solution to react to obtain the activated substrate.

[0007] According to some embodiments of the present application, the initiator is selected from one or more of (3-mercaptopropyl)trimethoxysilane, tetramethoxysilane-1-thiol, trimethoxysilane-1,1-dithiol, or trimethoxysilane-1,1,1-trithiol.

[0008] According to some embodiments of the present application, the solvent used in the initiator solution is selected from one or more of methanol, ethanol, toluene, tetrahydrofuran, or acetone.

[0009] According to some embodiments of the present application, the mass fraction of the initiator in the initiator solution is 1wt%-10wt%,

[0010] According to some embodiments of the present application, the catalyst includes an aqueous solution of hydrochloric acid; the molar concentration of the aqueous solution of hydrochloric acid is 0.01-0.2M, and the mass fraction in the mixed solution is 0.1wt%-2wt%.

[0011] According to some embodiments of the present application, the polymerizable monomer includes a phosphorylcholine compound; the auxiliary agent includes one or more of benzophenone, Irgacure series, Darocure series, Esacure series, and Lucirin series, and one or more of anhydrous methanol, ethanol, tetrahydrofuran, and acetone.

[0012] According to some embodiments of the present application, the polymerizable monomer includes 2-methacryloyloxyethyl phosphorylcholine.

[0013] According to some embodiments of the present application, the protective operation includes: introducing a first protective gas into the reaction container and freezing the reaction container to solidify the material in the reaction container; evacuating the reaction container, introducing a second protective gas, and thawing the solidified material.

[0014] On the other hand, the present application discloses a medical device having the above-mentioned anticoagulant coating.

[0015] The preparation method of the anticoagulation coating disclosed in the present application is to first coat a small molecule initiator coating on the surface of the substrate, and then directly polymerize the high molecular weight monomer in situ on the surface of the initiator coating. Since the steric hindrance of the small molecule initiator is much smaller than the polymer monomer used for grafting, the grafting density of the polymer monomer on the substrate surface is increased, thereby improving the anticoagulation effect of the anticoagulation coating. At the same time, not using a polymer as a primer further reduces the thickness of the anticoagulation coating and reduces the influence of the coating on other properties of the product itself.

[0016] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The present application will be further described in the form of exemplary embodiments, which will be described in detail by way of the accompanying drawings. These embodiments are not restrictive, and in these embodiments, the same number represents the same structure, wherein:

[0018] Figure 1 is an exemplary flow chart of a method for preparing an anticoagulation coating according to some embodiments of the present application;

[0019] Figure 2 is an exemplary schematic diagram of a method for preparing an anticoagulation coating according to some embodiments of the present application;

[0020] Figure 3 The anticoagulation effect of the test sample provided with the anticoagulation coating according to some embodiments of the present application is shown;

[0021] Figure 4 The anticoagulation effect of a comparative sample without the anticoagulation coating shown in the present application is shown. DETAILED DESCRIPTION

[0022] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below. In the following description, many specific details are set forth to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present application, so the present application is not limited by the specific embodiments disclosed below.

[0023] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as those generally understood by those skilled in the art to which this application belongs. The terms used in this application and in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application. Similar words such as "including" or "comprising" used in this application mean that the elements or objects appearing before the word cover the elements or objects listed after the word and their equivalents, without excluding other elements or objects. The terms "and / or" or "and / or" used in this application include any and all combinations of one or more related listed items. The terms "first", "second", etc. are only used to distinguish descriptions and cannot be understood as indicating or implying relative importance.

[0024] Flowcharts are used in the present application to illustrate the operations performed by the system according to the embodiments of the present application. It should be understood that the preceding or following operations are not necessarily performed accurately in order. On the contrary, various steps may be processed in reverse order or simultaneously. At the same time, other operations may also be added to these processes, or a certain step or several steps of operations may be removed from these processes.

[0025] Some preferred embodiments of the present application are described below. It should be noted that the following description is for the purpose of illustration and is not intended to limit the scope of protection of the present application. The steps involved in the present application can be performed accurately in sequence, or various steps can be processed in reverse order or simultaneously. At the same time, other operations can also be added to these processes, or one or more operations can be removed from these processes.

[0026] Figure 1 is an exemplary flow chart of a method for preparing an anticoagulation coating according to some embodiments of the present application. Figure 1 As shown, process 100 may include the following steps.

[0027] Step 110, coating the hydroxylated substrate with an initiator coating to obtain an activated substrate.

[0028] In some embodiments, the substrate may be formed by the surface of any suitable blood-contacting medical device, including intravascular interventional devices, extracorporeal circulation equipment, cardiovascular implants, blood-related devices, etc. or other related medical devices. Exemplary intravascular interventional devices may include vascular stents (e.g., coronary stents, carotid stents, peripheral vascular stents, aortic stents, etc.), catheters (e.g., central venous catheters, PICC catheters, angiography catheters, balloon dilatation catheters, guide catheters, microcatheters, etc.), guidewires (e.g., angiography guidewires, interventional surgery guidewires, hydrophilic coating guidewires, etc.). Exemplary extracorporeal circulation equipment may include hemodialysis-related equipment such as dialyzers, hemodialysis circuits, arteriovenous puncture needles, hemofilters, etc., or ECMO systems such as CMO circuits, oxygenators, centrifugal pumps, heat exchangers, etc. Exemplary cardiovascular implants may include artificial heart valves such as mechanical valves, bioprosthetic valves, transcatheter valves, etc., or other heart-related devices such as pacemaker wires, defibrillator wires, atrial occluders, ventricular assist devices, etc. Exemplary blood-related devices may include blood collection equipment such as blood collection needles, blood collection tubes, blood cell separators, platelet separators, etc. or blood transfusion equipment such as blood transfusion devices, blood bags, blood transfusion lines, blood warmers, etc. Other related medical devices may be monitoring equipment such as blood oxygen probes, blood pressure catheters, blood glucose monitoring needles, blood gas analysis catheters, etc.

[0029] In some embodiments, the hydroxylation treatment of the substrate can be performed using a chemical treatment method. For example, the surface hydroxylation can be completed by acid-base treatment of the substrate, such as using a strong acid (H2SO4, HNO3, etc.) or a strong base (NaOH, KOH, etc.) or a mixed treatment. For another example, the surface hydroxylation can be completed by oxidation treatment of the substrate, such as H2O2 oxidation, KMnO4 oxidation, potassium dichromate oxidation, Piranha solution treatment, etc. For another example, the surface hydroxylation of the substrate can be completed by a hydrolysis reaction, such as ester hydrolysis, amide hydrolysis, silane hydrolysis, etc. Taking Piranha solution treatment as an example, a fume hood can be used as a specific operating environment. 30% hydrogen peroxide is slowly added to concentrated sulfuric acid (volume ratio 7:3) to configure Piranha solution. The cleaned substrate is then completely immersed in the solution for 1-30 minutes until no bubbles are generated on the surface of the substrate. After that, the substrate is taken out and rinsed with purified water 3-6 times, and the surface hydroxylated substrate can be obtained after drying with an inert gas.

[0030] In some embodiments, the hydroxylation treatment of the substrate can also be carried out by physical treatment. For example, plasma treatment such as oxygen plasma, water vapor plasma, air plasma, etc. can be used. For another example, UV irradiation such as UV / O3 treatment, UV / H2O2 treatment, etc. can be used. For another example, corona treatment such as air corona, oxygen corona, etc. can be used. Taking plasma treatment as an example, the substrate can be suspended in a plasma treatment reactor, and the reactor is evacuated to -12Pa and oxygen is added to an appropriate positive pressure such as 50-65Pa and then maintained. The plasma treatment reactor is subsequently started to perform plasma surface activation treatment for 10 to 30 minutes to hydroxylate the substrate surface. Taking UV / O3 treatment as an example, the cleaned substrate can be placed in a preheated ultraviolet ozone (UVO) reactor and reacted for 10 to 60 minutes before taking out to obtain a surface hydroxylated substrate.

[0031] In some embodiments, the hydroxylation treatment of the substrate can also be performed using the aforementioned physical and chemical composite method. For example, it can be performed by plasma combined with chemical treatment, by UVO combined with chemical oxidation, etc. Alternatively, it can also be performed using a biological method. For example, it can be performed by enzyme catalysis such as lipase or hydrolase. This does not constitute a limitation of the present application.

[0032] In some embodiments, the hydroxylation of the substrate can be performed according to the material of the substrate. Exemplarily, when the substrate-related device is made of metal materials, for example, vascular stents such as self-expanding stents, mechanical heart valves, atrial occluders, blood flow guide devices, embolic protection umbrellas, guide wires, etc., the hydroxylation treatment can be carried out by acid-base treatment, anodizing, plasma treatment, etc. When the substrate-related device is made of polymer materials, for example, catheters, drainage tubes, vascular stents such as self-expanding stents, blood flow guide devices, atrial occluders, embolic protection umbrellas, etc., the hydroxylation treatment can be carried out by plasma treatment, UVO, chemical oxidation, etc. When the substrate-related device is made of ceramic materials, for example, artificial joints, metal stent ceramic coatings, blood filter ceramic membranes, etc., the hydroxylation treatment can be carried out by acid-base treatment, hydrothermal treatment, plasma treatment, etc.

[0033] The above substrate for surface hydroxylation may be pre-cleaned. For example, the substrate may be immersed in an organic solvent such as methanol, ethanol, isopropanol, acetone, methyl ethyl ketone, n-hexane, petroleum ether, tetrahydrofuran, cyclohexane, deionized water, distilled water, ultrapure water, etc., and ultrasonicated for 5-60 minutes. After completion, place it on dust-free paper to dry. Alternatively, the substrate may be purged with an inert gas such as nitrogen or argon, or an active gas such as ozone or oxygen plasma to achieve surface cleaning.

[0034] In certain embodiments, the coating of the initiator coating can be carried out based on an initiator. Exemplary, the initiator can be provided to configure an initiator solution. The initiator can be selected from an organic compound with methoxy silicon and / or sulfhydryl. When the initiator is a compound comprising methoxy silicon, it can contain other active functional groups at the same time, including but not limited to hydroxyl, amino, amide, aldehyde etc. Illustrative but non-limiting examples of such initiators include 3-hydroxypropyltrimethoxysilane, 2-hydroxyethyltrimethoxysilane, 2-hydroxypropyltrimethoxysilane, 4-hydroxybutyltrimethoxysilane, 1-hydroxy-2-methylethyltrimethoxysilane, 3-hydroxy-4-methylbutyltrimethoxysilane, 5-hydroxypentyltrimethoxysilane, 2,3-dihydroxypropyltrimethoxysilane, 3-aminopropyltrimethoxysilane, 2-aminoethyltrimethoxysilane, 4-aminobutyltrimethoxysilane, 2-aminopropyltrimethoxysilane, N-methyl-3-aminopropyltrimethoxysilane, N-ethyl-2-aminoethyltrimethoxysilane, N,N-dimethyl-3-aminopropyltrimethoxysilane, 2-(2-aminoethyl)aminoethyltrimethoxysilane, Silane, N-acetyl-3-aminopropyltrimethoxysilane, N-acetyl-2-aminoethyltrimethoxysilane, N-propionyl-3-aminopropyltrimethoxysilane, N-methyl-2-amidoethyltrimethoxysilane, amidomethyltrimethoxysilane, 3-amidopropyltrimethoxysilane, N-acetyl-4-aminobutyltrimethoxysilane, N-ethyl-2-amidoethyltrimethoxysilane, 3-aldehydepropyltrimethoxysilane, 2-aldehydeethyltrimethoxysilane, aldehydemethyltrimethoxysilane, 4-aldehydebutyltrimethoxysilane, 2-aldehydepropyltrimethoxysilane, aldehydemethyldimethoxymethylsilane, 5-aldehydepentyltrimethoxysilane, 2-aldehydepropyltrimethoxysilane, 3-aldehyde-4-methylbutyltrimethoxysilane, etc.

[0035] When the initiator has a mercapto group, it may also have other active functional groups that can react with a hydroxyl group. Exemplarily, the functional group may be an isocyanate group, and the initiator may include 2-mercaptoethyl isocyanate, 3-mercaptopropyl isocyanate, 2-methyl-2-mercaptoethyl isocyanate, 4-mercaptobutyl isocyanate, 4-mercaptomethylphenyl isocyanate, etc. The functional group may also be an epoxy group, and the initiator may include 2-mercaptoethoxy oxirane, 2-epoxypropoxy-3-mercaptobutane, 3-mercaptopropoxy oxirane, 4-mercaptomethylphenoxy oxirane, 2-mercaptoethoxymethyl oxirane, etc. The functional group may also be an anhydride group, and the initiator may include mercaptoacetic anhydride, 2-mercaptoethyl acetic anhydride, 4-mercaptomethylbenzoyl acetic anhydride, 3-mercaptopropyl propionic anhydride, 2-methyl-2-mercaptoacetic anhydride, etc. The functional group may also be an acyl chloride group, and the initiator may include mercaptoacetyl chloride, 2-mercaptoacetyl chloride, 2-methyl-2-mercaptopropionyl chloride, 3-mercaptopropionyl chloride, 4-mercaptomethylbenzoyl chloride, etc. The functional group may also be a haloalkyl group, and the initiator may include 3-mercapto-1-chloropropane, 1-mercapto-2-chloromethylpropane, 5-mercapto-1-chloropentane, 4-mercaptomethylbenzyl chloride, 1-mercapto-2-methyl-3-chloropropane, etc. The functional group may also be an ester group, and the initiator may include methyl thioacetate, methyl 2-mercaptopropionate, methyl 2-methyl-2-mercaptopropionate, ethyl 3-mercaptobutyrate, methyl 4-mercaptomethylbenzoate, etc.

[0036] The initiator may also include methoxysilane and mercapto. Exemplarily, the initiator may include but is not limited to 3-(mercaptopropyl)trimethoxysilane, 2-(mercaptoethyl)trimethoxysilane, 3-(mercaptopropyl)dimethoxymethylsilane, 4-(mercaptobutyl)trimethoxysilane, 2-methyl-2-(mercaptoethyl)trimethoxysilane, 2,2-dimethyl-2-(mercaptoethyl)trimethoxysilane, 2-ethyl-2-(mercaptoethyl)trimethoxysilane, 2-methyl The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group. The initiator may be a methoxysilane or a sulfhydryl group.

[0037] In some embodiments, solvents such as n-hexane, cyclohexane, petroleum ether, toluene, xylene, tetrahydrofuran, ether, dioxane, methyl tert-butyl ether (MTBE), acetone, methyl ethyl ketone (MEK), cyclohexanone, methyl isobutyl ketone (MIBK), ethyl acetate, dimethyl carbonate, N, N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAc), N-methylpyrrolidone (NMP), pyridine, methanol, ethanol, isopropanol, n-butanol, trifluoroacetic acid, dichloromethane, chloroform, dimethyl sulfoxide (DMSO), acetonitrile, nitromethane, etc. can be used to dissolve the initiator to obtain the initiator solution. In some embodiments, the mass fraction of the initiator in the initiator solution can be 1wt%-10wt%. For example, 1wt%, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, 10wt%. Alternatively, the mass fraction of the initiator in the initiator solution may be any value within the above numerical range.

[0038] After obtaining the initiator solution, a catalyst can be added thereto to obtain a mixed solution for activating the substrate after hydroxylation. The catalyst can be an acid solution, including but not limited to sulfuric acid solution, hydrochloric acid solution, nitric acid solution, perchloric acid, hydrobromic acid, hydroiodide acid, phosphoric acid, sulfurous acid, hydrofluoric acid, nitrous acid, hypochlorous acid, chlorous acid, phosphorous acid, cyanic acid, boric acid, etc. or any combination thereof. Alternatively or preferably, the catalyst can be a hydrochloric acid solution, and its amount of substance concentration can be between 0.01M-0.2M, for example, 0.01M, 0.02M, 0.03M, 0.04M, 0.05M, 0.06M, 0.07M, 0.08M, 0.09M, 0.1M, 0.11M, 0.12M, 0.13M, 0.14M, 0.15M, 0.16M, 0.17M, 0.18M, ​​0.19M, 0.2M, etc. Alternatively, the amount of substance concentration of the hydrochloric acid solution may be 0.05M. The mass fraction of the hydrochloric acid solution in the initiator solution may be 0.1wt%-2wt%. Alternatively or preferably, the mass fraction of the hydrochloric acid solution in the initiator solution may be 0.2wt%-1.8wt%. Alternatively or preferably, the mass fraction of the hydrochloric acid solution in the initiator solution may be 0.5wt%-1.5wt%. Alternatively or preferably, the mass fraction of the hydrochloric acid solution in the initiator solution may be 0.7wt%-1.3wt%. Alternatively or preferably, the mass fraction of the hydrochloric acid solution in the initiator solution may be 0.9wt%-1.1wt%. Alternatively or preferably, the mass fraction of the hydrochloric acid solution in the initiator solution may be 0.1wt%, 1wt%, 2wt%.

[0039] In some embodiments, the hydroxylated substrate can be placed in the mixed solution to obtain the activated substrate. Any operation method that can accelerate the reaction rate, including but not limited to temperature regulation such as heating / keeping warm, pressure regulation such as increasing the reaction pressure or using a pressure reactor, ultrasonic dispersion, illumination, electric field application, stirring, etc. can be applied to the process. A non-limiting example can be to completely immerse the hydroxylated substrate in the mixed solution and soak it at a temperature of 30°C-50°C (for example, 40°C) for 1-24 hours.

[0040] In some embodiments, the activated substrate may be subjected to necessary cleaning treatment before subsequent operations, for example, the activated substrate taken out from the mixed solution may be rinsed and cleaned using a solvent such as methanol, ethanol, toluene, tetrahydrofuran, acetone, deionized water, or the like.

[0041] Step 120, placing the activated substrate, polymerizable monomers and auxiliary agents in a reaction container, and adjusting the interior of the reaction container to a water-free and oxygen-free environment in one or more protection operations.

[0042] In some embodiments, the polymerizable monomers can be used to form an anticoagulant coating, including but not limited to high molecular weight polymers such as polyethylene glycol (PEG) and its derivatives, polytetrafluoroethylene (PTFE), polydimethylsiloxane (PDMS), polyvinyl pyrrolidone (PVP), polyacrylic acid and its esters, polyurethane (PU), polyvinyl alcohol (PVA), polymethyl methacrylate (PMMA), etc., phospholipid compounds such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), dipalmitoylphosphatidylcholine (DPPC), etc., heparin and its derivatives such as heparin sulfonate, low molecular weight heparin, heparin compounds, etc., zwitterionic compounds such as betaine derivatives, sulfobetaine, carboxybetaine, etc., biomolecules such as albumin, gelatin, hyaluronic acid, chitosan, alginate, etc., functional monomers such as acrylamide, methacrylate, vinyl pyrrolidone, vinyl alcohol, etc. or any combination thereof.

[0043] In combination with the above description, the polymerized monomer can react with the initiator coating on the activated substrate. Based on this, the polymerized monomer can include a compound that can react with the above-mentioned initiator. Therefore, the polymerized monomer can include a compound having a functional group that can react with a hydroxyl, an amino, an amide, an aldehyde, or a sulfhydryl. In some embodiments, the initiator has methoxysilane and a sulfhydryl group at the same time, and the methoxysilane reacts with the substrate with hydroxylation on the surface to form an initiator coating, and the sulfhydryl group can react with the polymerized monomer to form an anticoagulant coating. Based on this, the polymerized monomer can be a compound that can react with a sulfhydryl group and can achieve an anticoagulant effect, such as preventing thrombosis. Exemplary, the polymerized monomer can include a phosphatidylcholine (PC) compound. Phosphatidylcholine compounds can include but are not limited to natural phosphatidylcholine compounds, synthetic phosphatidylcholine compounds, modified phosphatidylcholine compounds, functionalized phosphatidylcholine compounds, etc. Some non-limiting examples of natural phosphatidylcholine compounds can be egg phosphatidylcholine (Egg-PC), soybean phosphatidylcholine (Soy-PC), brain phosphatidylcholine (Brain-PC), cardiolipin (Heart-PC), etc. Some non-limiting examples of synthetic phosphatidylcholine compounds can be dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), 1-stearoyl-2-oleoylphosphatidylcholine (SOPC), didecanoylphosphatidylcholine (DLPC), dilauroylphosphatidylcholine (DLPC), etc. Some non-limiting examples of modified phosphatidylcholine compounds can be 2-methacryloyloxyethylphosphatidylcholine (MPC), 2-acryloyloxyethylphosphatidylcholine (APC), methacrylate-2-phosphatidylcholine ethyl ester (MPCE), poly(2-methacryloyloxyethylphosphatidylcholine) (Poly(MPC)), poly(MPC-co-BMA) (copolymer of MPC and butyl methacrylate), poly(MPC-co-HEMA) (copolymer of MPC and 2-hydroxyethyl methacrylate), etc.Some non-limiting examples of functionalized phosphatidylcholine compounds can be thiol-functionalized phosphatidylcholine (e.g., mercaptoethyl phosphatidylcholine, distearoyl-mercaptoethyl phosphatidylcholine, etc.), amino-functionalized phosphatidylcholine (e.g., aminoethyl phosphatidylcholine, dipalmitoyl-aminoethyl phosphatidylcholine, etc.), carboxyl-functionalized phosphatidylcholine (e.g., carboxyethyl phosphatidylcholine), biotin-labeled phosphatidylcholine (e.g., biotin-phosphatidylcholine conjugate, dipalmitoyl-biotin phosphatidylcholine, etc.), fluorescently labeled phosphatidylcholine (e.g., NBD-labeled phosphatidylcholine, rhodamine-labeled phosphatidylcholine, etc.), photocross-linked phosphatidylcholine (e.g., benzoyl phosphatidylcholine, distearoyl-azo phosphatidylcholine, etc.), polymerized phosphatidylcholine (e.g., PEGylated phosphatidylcholine, PLGA-phosphatidylcholine copolymer, etc.). In some embodiments, the phosphatidylcholine compound may include 2-methacryloyloxyethyl phosphatidylcholine.

[0044] In some embodiments, the auxiliary agent can be used as a reaction initiating component for reacting (or grafting) with the polymerized monomer in the initiator coating or a solvent for providing a reaction site. Exemplarily, the auxiliary agent may include an initiator and a reaction solvent. The initiator may include a photoinitiator (for example, diphenylacetophenone, 2,2-dimethoxy-2-phenylacetophenone (DMPA), 2,2-diethoxyacetophenone, phenacyl, benzophenone, 2,4-dihydroxybenzophenone, Irgacure series (such as Irgacure 184, Irgacure 651, etc.), Darocure series (such as Darocure 1173, Darocure 2959, etc.), Esacure series (such as Esacure KIP 100, Esacure KIP 150, etc.), Lucirin series (such as Lucirin TPO, Lucirin DP, etc.), azo initiators (for example, azobisisobutyronitrile AIBN, azodimethylbutyronitrile AMBN, 2,2'-azobis(2-methylpropionamide), 2,2'-azobis(2,4-dimethylvaleronitrile, etc.), peroxide compounds (for example, benzoyl peroxide BPO, tert-butyl peroxybenzoate TBPB, di-tert-butyl peroxide DTBP, cumyl peroxide CHP, etc.), redox systems (for example, ammonium persulfate / TEMED, potassium persulfate / ascorbic acid, potassium persulfate / sodium sulfite, etc.). The reaction solvent may include methanol, ethanol, isopropanol, acetone, acetonitrile, dimethylsulfamide, chloroform, etc. The reaction solvent may also be treated to remove water. The above initiating catalyst and water removal agent, etc. may be adjusted according to actual conditions and are not specifically limited.

[0045] In some embodiments, the protection operation may include the following steps. Step 1, introducing a protective gas into the reaction vessel and freezing it to solidify the material in the reaction vessel. The protective gas may be nitrogen, argon, helium, carbon dioxide, etc., and the reaction vessel may be a multi-mouth flask, or a flask connected to a Schlenk tube. Freezing may be achieved by an ice bath, a brine ice bath, a dry ice bath, liquid nitrogen, a freezer, a circulating refrigerator, etc. Step 2, introducing a protective gas after evacuating the reaction vessel, and thawing the solidified material. Exemplarily, one port in a two-mouth flask is used to introduce a protective gas, and the other port is used to evacuate. Alternatively, the switching of the vacuum environment and the protective environment of the flask in the Schlenk tube system is achieved by a Schlenk operation. The protective gas used in step 2 may be the same or different from the protective gas in step 1. For example, both may be nitrogen, or nitrogen or helium, respectively. This is not restrictive. Thawing may be performed by a warm water bath, for example, by placing the reaction vessel in room temperature water. By repeatedly performing the above protection operation multiple times, for example, twice, three times, four times, etc., the interior of the reaction container can be adjusted to a water-free and oxygen-free environment.

[0046] Step 130, placing the reaction container under target reaction conditions to react so that the polymerizable monomers are polymerized in situ on the activated substrate, thereby obtaining the anticoagulation coating.

[0047] In some embodiments, the target reaction conditions can be determined according to the selection of the aforementioned initiator, polymerization monomer and initiator. Exemplarily, when the initiator is a compound containing both methoxysilane and thiol, such as 3-(mercaptopropyl) trimethoxysilane, 4-(mercaptobutyl) trimethoxysilane, (3-trimethoxysilylpropyl) dithiol, (3-trimethoxysilylpropyl) trithiol, etc., and the polymerization monomer is a phosphatidylcholine compound such as 2-methacryloyloxyethyl phosphorylcholine (MPC), when the initiator is a photoinitiator, the target reaction conditions can be ultraviolet light conditions, for example, the reaction vessel is placed in a dark box and provided with ultraviolet light conditions by an ultraviolet lamp installed on the inner wall, and the wavelength can be 300nm-365nm, or 340nm-360nm. When the initiator is an azo compound such as AIBN, the target reaction conditions can be constant temperature maintenance after heating, for example, a reaction temperature of 60°C-65°C is provided by an oil bath. When the initiator is a redox system such as ammonium persulfate / TEMED, the target reaction conditions can be room temperature conditions, for example, directly reacting in a fume hood or on a test bench. The target reaction conditions can also include stirring, ventilation, etc. Exemplarily, a magnet can be placed in the reaction vessel, and the reaction can be placed on a magnetic stirrer. Equipment such as a blower can be used to maintain gas exchange in the environment of the reaction vessel.

[0048] During the reaction, the polymerized monomers can react with the initiator to complete the attachment to the substrate. At the same time, a polymerization process occurs between the polymerized monomers, and a polymerization reaction is directly carried out at a specific position / interface (i.e., the substrate surface). This does not require the pre-synthesis of polymer chains and linking to the substrate surface, effectively reducing the steric hindrance during the synthesis of the anticoagulant coating and increasing the grafting density of the polymerized monomers on the substrate.

[0049] After a period of reaction, for example, after 0.5 hours to 20 hours, the substrate with the anticoagulant coating can be taken out from the reaction vessel and extracted multiple times in a solvent such as a polar solvent (including but not limited to water, methanol, ethanol, isopropanol, n-butanol, acetone, methyl ethyl ketone, cyclohexanone, tetrahydrofuran, dioxane, ethyl acetate, acetonitrile, N,N-dimethylformamide, dichloromethane, chloroform, etc.) to wash away the unreacted polymerized monomers and the auxiliary agent attached to the surface. The substrate can then be placed in an oven for heating and annealing. After the end, it is immersed in a solvent such as a polar solvent (for example, various solvents as described above) and ultrasonicated for a period of time. The solvent is replaced and the ultrasonication is repeated multiple times to obtain the final substrate with an anticoagulant coating.

[0050] Figure 2 An exemplary schematic diagram of a method for preparing an anticoagulation coating according to some embodiments of the present application is shown, as shown in FIG. Figure 2 As shown, the substrate is prepared from a nickel-titanium alloy. The nickel-titanium alloy treated with surface hydroxylation reacts with an initiator (3-mercaptopropyl) trimethoxysilane in a methanol solvent under heating conditions to form an initiator coating on its surface. Subsequently, a polymerizable monomer 2-methacryloyloxyethyl phosphorylcholine (MPC) used to form an anticoagulant coating is directly in situ polymerized on the initiator coating to form an anticoagulant coating on the surface of the nickel-titanium alloy.

[0051] The preparation method of the anticoagulant coating disclosed in the present application is that the polymer material for anticoagulant effect is directly polymerized in situ on the surface of the device to form the anticoagulant coating. Not using a polymer primer not only reduces the total thickness of the coating, but also avoids the low grafting efficiency caused by steric hindrance during in situ polymerization.

[0052] On the other hand, the present application discloses a medical device having an anticoagulant coating as described above. The medical device may include any suitable blood-contact medical device, including but not limited to intravascular interventional devices, extracorporeal circulation equipment, cardiovascular implants, blood-related devices, etc. or other related medical devices. Exemplary intravascular interventional devices may include vascular stents (e.g., coronary stents, carotid stents, peripheral vascular stents, aortic stents, etc.), catheters (e.g., central venous catheters, PICC catheters, angiographic catheters, balloon dilatation catheters, guide catheters, microcatheters, etc.), guide wires (e.g., angiographic guide wires, interventional surgical guide wires, hydrophilic coating guide wires, etc.). Exemplary extracorporeal circulation equipment may include hemodialysis-related equipment such as dialyzers, hemodialysis circuits, arteriovenous puncture needles, blood filters, etc., or ECMO systems such as CMO circuits, oxygenators, centrifugal pumps, heat exchangers, etc. Exemplary cardiovascular implants may include artificial heart valves such as mechanical valves, bioprosthetic valves, transcatheter valves, etc., or other heart-related devices such as pacemaker wires, defibrillator wires, atrial occluders, ventricular assist devices, etc. Exemplary blood-related devices may include blood collection equipment such as blood collection needles, blood collection tubes, blood cell separators, platelet separators, etc. or blood transfusion equipment such as blood transfusion devices, blood bags, blood transfusion lines, blood warmers, etc. Other related medical devices may be monitoring devices such as blood oxygen probes, blood pressure catheters, blood glucose monitoring needles, blood gas analysis catheters, etc. The medical device disclosed in the present application has an anticoagulant coating with a high-efficiency anticoagulant function.

[0053] The present application is further described in detail below in conjunction with the examples. It should be noted that the following examples are only used to illustrate the present application and are not used to limit the scope of protection claimed in the present application.

[0054] Example 1 - Preparation of anticoagulation coating

[0055] 1) Material cleaning:

[0056] Clean the surface of the blood flow guide device (made of nickel-titanium / cobalt-chromium alloy, referred to as the product in the following). Immerse the product completely in ethanol solvent and ultrasonicate for 5-60 minutes. After cleaning, place the product on dust-free paper to dry for later use

[0057] 2) Surface pretreatment:

[0058] Slowly add 30% hydrogen peroxide to concentrated sulfuric acid in a volume ratio of 7:3. Immerse the cleaned product completely in the piranha solution for 1-30 minutes until no bubbles are generated on the surface of the material. Then take out the sample with tweezers, rinse it with purified water 3-6 times, and blow dry it with inert gas;

[0059] 3) Coating initiator:

[0060] The initiator is prepared into a 1wt% solution. Here, the initiator is (3-mercaptopropyl)trimethoxysilane. The solvent used is methanol. Then, 0.1wt% 0.05M hydrochloric acid aqueous solution is added to the prepared solution. Then, the surface pretreated product is completely immersed in it and soaked at 40 degrees Celsius for 1-24 hours.

[0061] 4) In-situ polymerization:

[0062] The product is placed in a dry Schlenk bottle, and the feed is calculated according to the surface area of ​​the product. 100 mg of 2-methacryloyloxyethyl phosphorylcholine, 1 mg of benzophenone, 1 mL of anhydrous methanol and a magnet are added per square centimeter. After the addition is completed, a protector is passed through, and liquid nitrogen is used to freeze the Schlenk bottle for 5-20 minutes until the liquid in the bottle solidifies. Then vacuum for 10-20 minutes. After that, the vacuum is turned off, the protective gas is passed through, and the Slenck bottle is placed in room temperature water to thaw. Repeat this step three times, and the Schlenk bottle will be an anhydrous and oxygen-free environment. The Schlenk bottle is placed in a UV reactor (340-360 nm), and a magnetic stirrer (100-400 rpm) is turned on, and the reaction is 0.5-20 hours.

[0063] 5) Post-processing and cleaning:

[0064] The sample was taken out of the Schlenk bottle and extracted in methanol for 3 times, and then placed in an oven and annealed at 80°C for 30-90 minutes. After annealing, the sample was completely immersed in acetone and ultrasonicated for 10 minutes, and the solvent was changed and ultrasonicated for 2-3 times.

[0065] Example 2 - Preparation of anticoagulation coating

[0066] 1) Material cleaning:

[0067] Clean the surface of the interventional catheter (made of high molecular weight polyester material, referred to as the product in the following). Immerse the product completely in ethanol solvent and ultrasonicate for 5-60 minutes. After cleaning, place the product on dust-free paper to dry for later use.

[0068] 2) Surface pretreatment:

[0069] After cleaning the product, put it into the preheated UVO reactor and take it out after reacting for 10-60 minutes;

[0070] 3) Coating initiator:

[0071] The initiator is prepared into a 10wt% solution. Here, the initiator is (3-mercaptopropyl)trimethoxysilane. The solvent used is methanol. Then, 2wt% 0.05M hydrochloric acid aqueous solution is added to the prepared solution. Then, the surface pretreated product is completely immersed in it and soaked at 40 degrees Celsius for 1-24 hours.

[0072] 4) In-situ polymerization:

[0073] The product is placed in a dry Schlenk bottle, and the feed is calculated according to the surface area of ​​the product. 100 mg of 2-methacryloyloxyethyl phosphorylcholine, 1 mg of benzophenone, 1 mL of anhydrous methanol and a magnet are added per square centimeter. After the addition is completed, a protector is passed through, and liquid nitrogen is used to freeze the Schlenk bottle for 5-20 minutes until the liquid in the bottle solidifies. Then vacuum for 10-20 minutes. After that, the vacuum is turned off, the protective gas is passed through, and the Slenck bottle is placed in room temperature water to thaw. Repeat this step three times, and the Schlenk bottle will be an anhydrous and oxygen-free environment. The Schlenk bottle is placed in a UV reactor (340-360 nm), and a magnetic stirrer (100-400 rpm) is turned on, and the reaction is 0.5-20 hours.

[0074] 5) Post-processing and cleaning:

[0075] The sample was taken out of the Schlenk bottle and extracted in methanol for 3 times, and then placed in an oven and annealed at 80°C for 30-90 minutes. After annealing, the sample was completely immersed in acetone and ultrasonicated for 10 minutes, and the solvent was changed and ultrasonicated for 2-3 times.

[0076] Chandler Loop In Vitro Anticoagulation Test

[0077] 1) Place the coated dense mesh stent (test product) and the bare stent of the same specification (control product) into the same silicone tube through a plastic rod;

[0078] 2) Inject 5 mL of purified water into the silicone tube;

[0079] 3) Adjust the water bath in the Chandlar Loop System to 37°C and install the silicone tubing on the device;

[0080] 4) Turn on the Chandlar Loop System and rotate for 10 minutes;

[0081] 5) Stop the rotation and pour out the purified water in the silicone tube;

[0082] 6) Add 5 mL of anticoagulated rabbit blood into a 20 mL vial, use a pipette to add 0-100 μL of 0.2 M CaCl2 solution into the rabbit blood, stir rapidly, and then add the calcified rabbit blood into the silicone tube placed in the dense mesh support;

[0083] 7) Start the rotation and continue for 1-30 minutes;

[0084] 8) Prepare 40 mL of 4 wt % glutaraldehyde aqueous solution, take two 20 mL vials, and fill each with 20 mL of 4 wt % glutaraldehyde aqueous solution;

[0085] 9) Stop the rotation, remove the silicone tube, pour out the rabbit blood, take out the two dense mesh stents with tweezers (or cut the silicone tube at the position of the dense mesh stent with scissors, keep the dense mesh stent in the silicone tube), and soak them in a vial filled with 4wt% glutaraldehyde aqueous solution for 30 minutes, and put the test sample and the control sample into two vials respectively;

[0086] 10) Take out the samples respectively and place them in ethanol solution for dehydration;

[0087] 11) Observe the formation of thrombus on the dense mesh stent and take photos for record.

[0088] The test results are as follows Figure 3 and Figure 4 As shown, Figure 3 The thrombosis of the test article is shown, and no thrombosis occurs on the stent. Figure 4 The thrombosis of the control is shown, and a large area of ​​thrombus is attached to the stent (such as Figure 4 The black circle in the middle). It can be seen that the anticoagulation coating disclosed in the present application has an excellent anticoagulation effect.

[0089] The present application has described the basic concepts. Obviously, for those skilled in the art, the above detailed disclosure is only for example and does not constitute a limitation of the present application. Although not explicitly stated herein, those skilled in the art may make various modifications, improvements and amendments to the present application. Such modifications, improvements and amendments are suggested in the present application, so such modifications, improvements and amendments still belong to the spirit and scope of the exemplary embodiments of the present application.

[0090] At the same time, the present application uses specific words to describe the embodiments of the present application. For example, "one embodiment", "an embodiment", and / or "some embodiments" refer to a certain feature, structure or characteristic related to at least one embodiment of the present application. Therefore, it should be emphasized and noted that "one embodiment" or "an embodiment" or "an alternative embodiment" mentioned twice or multiple times in different positions in the present application does not necessarily refer to the same embodiment. In addition, some features, structures or characteristics in one or more embodiments of the present application can be appropriately combined.

[0091] Similarly, it should be noted that in order to simplify the description of the disclosure of this application and thus help understand one or more embodiments of the invention, in the above description of the embodiments of this application, multiple features are sometimes combined into one embodiment or its description. However, this disclosure method does not mean that the features required by the object of this application are more than the features mentioned in the claims. In fact, the features of the embodiments are less than all the features of the single embodiment disclosed above.

[0092] Finally, it should be understood that the embodiments described in this application are only used to illustrate the principles of the embodiments of the present application. Other variations may also fall within the scope of the present application. Therefore, as an example and not a limitation, the alternative configurations of the embodiments of the present application may be considered to be consistent with the teachings of the present application. Accordingly, the embodiments of the present application are not limited to the embodiments explicitly introduced and described in the present application.

Claims

1. A method for preparing an anticoagulation coating, characterized in that: The preparation method comprises: applying an initiator coating to the hydroxylated substrate to obtain an activated substrate; Placing the activated substrate, polymerizable monomers and auxiliary agents in a reaction container, and adjusting the interior of the reaction container to an anhydrous and oxygen-free environment during one or more protection operations; The reaction container is placed under target reaction conditions to react so that the polymerizable monomers are polymerized in situ on the activated substrate, thereby obtaining the anticoagulation coating.

2. The preparation method according to claim 1, characterized in that: The step of obtaining an activated substrate comprises: Providing an initiator and preparing an initiator solution; the initiator is selected from compounds having methoxysilane and / or mercapto groups; A catalyst is added to the initiator solution to obtain a mixed solution, and the hydroxylated substrate is placed in the mixed solution for reaction to obtain the activated substrate.

3. The preparation method according to claim 2, characterized in that: The initiator is selected from one or more of (3-mercaptopropyl)trimethoxysilane, tetramethoxysilane-1-thiol, trimethoxysilane-1,1-dithiol, and trimethoxysilane-1,1,1-trithiol.

4. The preparation method according to claim 2, characterized in that: The solvent used in the initiator solution is selected from one or more of methanol, ethanol, toluene, tetrahydrofuran, or acetone.

5. The preparation method according to claim 2, characterized in that: The mass fraction of the initiator in the initiator solution is 1 wt%-10 wt%.

6. The preparation method according to claim 2, characterized in that: The catalyst comprises a hydrochloric acid aqueous solution; the molar concentration of the hydrochloric acid aqueous solution is 0.01-0.2M, and the mass fraction of the hydrochloric acid aqueous solution in the mixed solution is 0.1wt%-2wt%.

7. The preparation method according to claim 1, characterized in that: The polymerizable monomers include phosphorylcholine compounds; the auxiliary agents include one or more of benzophenone, Irgacure series, Darocure series, Esacure series, and Lucirin series, and one or more of anhydrous methanol, ethanol, tetrahydrofuran, and acetone.

8. The preparation method according to claim 7, characterized in that: The polymerizable monomer includes 2-methacryloyloxyethyl phosphorylcholine.

9. The preparation method according to claim 1, characterized in that: The protection operation includes: After introducing a first protective gas into the reaction container, the reaction container is frozen to solidify the material in the reaction container; After the reaction container is evacuated, a second protective gas is introduced, and the solidified material is thawed.

10. A medical device having an anticoagulation coating according to any one of claims 1 to 9.

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